Difference between revisions of "Team:NCTU Formosa/Notebook"

(Prototype team page)
 
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{{NCTU_Formosa}}
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{{NCTU_Formosa/test}}
 
<html>
 
<html>
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<head>
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<script src="https://ajax.googleapis.com/ajax/libs/jquery/3.1.0/jquery.min.js"></script>
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<link href="https://2016.igem.org/Team:NCTU_Formosa/wiki_reset?action=raw&ctype=text/css" rel="stylesheet">
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<h1>Notebook</h1>
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html, body, div, span, applet, object, iframe,
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<p> Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.</p>
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h1, h2, h3, h4, h5, h6, p, blockquote, pre,
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<div class="clear"></div>
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-moz-box-sizing: border-box;
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<div class="column two_thirds_size">
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*:after {
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<h3>What should this page have?</h3>
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margin: 0;
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<ul>
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padding: 0;
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<li>Chronological notes of what your team is doing.</li>
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  font-family: 'Open Sans', sans-serif;
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<li> Brief descriptions of daily important events.</li>
+
  font-weight: 400;
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<li>Pictures of your progress. </li>
+
}
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<li>Mention who participated in what task.</li>
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−
</ul>
+
  
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 +
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 +
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 +
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 +
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 +
 +
</style>
 +
</head>
 +
<body>
 +
    <div class="img-container">
 +
        <img src="https://static.igem.org/mediawiki/2018/3/31/T--NCTU_Formosa--Notebook_cover.png" width="100%">
 +
    </div>
 +
<!-- Contenedor principal -->
 +
<div style="margin:auto;position:relative;">
 +
<h1>Wet Lab</h1>
 
</div>
 
</div>
 +
<div class="menu">
  
−
<div class="column third_size">
 
−
<div class="highlight decoration_A_full">
 
−
<h3>Inspiration</h3>
 
−
<p>You can see what others teams have done to organize their notes:</p>
 
  
−
<ul>  
+
<ul class="menu__tabs">
−
<li><a href="https://2014.igem.org/Team:ATOMS-Turkiye/Notebook">2014 ATOMS-Turkiye</a></li>
+
<li><a class="active" href="#item-1">Jul</a></li>
−
<li><a href="https://2014.igem.org/Team:Tec-Monterrey/ITESM14_project.html#tab_notebook">2014 Tec Monterrey</a></li>
+
<li><a href="#item-2">Aug</a></li>
−
<li><a href="https://2014.igem.org/Team:Kyoto/Notebook/Magnetosome_Formation#title">2014 Kyoto</a></li>
+
<li><a href="#item-3">Sep</a></li>
−
<li><a href="https://2014.igem.org/Team:Cornell/notebook">2014 Cornell</a></li>
+
<li><a href="#item-4">Oct</a></li>
 +
</ul>
 +
<section class="menu__wrapper">
 +
 
 +
<article id="item-1" class="menu__item item-active">
 +
<table style="width:100%" class="calender">
 +
                    <tr>
 +
                        <th>Sun</th>
 +
                        <th>Mon</th>
 +
                        <th>Tue</th>
 +
                        <th>Wed</th>
 +
                        <th>Thu</th>
 +
                        <th>Fri</th>
 +
                        <th>Sat</th>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item td-active"><a href="#July-1">1</a></td>
 +
                        <td class="td-item"><a href="#July-2">2</a></td>
 +
                        <td class="td-item"><a href="#July-3">3</a></td>
 +
                        <td class="td-item"><a href="#July-4">4</a></td>
 +
                        <td class="td-item"><a href="#July-5">5</a></td>
 +
                        <td class="td-item"><a href="#July-6">6</a></td>
 +
                        <td class="td-item"><a href="#July-7">7</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#July-8">8</a></td>
 +
                        <td class="td-item"><a href="#July-9">9</a></td>
 +
                        <td class="td-item"><a href="#July-10">10</a></td>
 +
                        <td class="td-item"><a href="#July-11">11</a></td>
 +
                        <td class="td-item"><a href="#July-12">12</a></td>
 +
                        <td class="td-item"><a href="#July-13">13</a></td>
 +
                        <td class="td-item"><a href="#July-14">14</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#July-15">15</a></td>
 +
                        <td class="td-item"><a href="#July-16">16</a></td>
 +
                        <td class="td-item"><a href="#July-17">17</a></td>
 +
                        <td class="td-item"><a href="#July-18">18</a></td>
 +
                        <td class="td-item"><a href="#July-19">19</a></td>
 +
                        <td class="td-item"><a href="#July-20">20</a></td>
 +
                        <td class="td-item"><a href="#July-21">21</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#July-22">22</a></td>
 +
                        <td class="td-item"><a href="#July-23">23</a></td>
 +
                        <td class="td-item"><a href="#July-24">24</a></td>
 +
                        <td class="td-item"><a href="#July-25">25</a></td>
 +
                        <td class="td-item"><a href="#July-26">26</a></td>
 +
                        <td class="td-item"><a href="#July-27">27</a></td>
 +
                        <td class="td-item"><a href="#July-28">28</a></td>
 +
                    </tr>
 +
                    <tr class="bottom"><!--calender bottom line-->
 +
                        <td class="td-item"><a href="#July-29">29</a></td>
 +
                        <td class="td-item"><a href="#July-30">30</a></td>
 +
                        <td class="td-item"><a href="#July-31">31</a></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                    </tr>
 +
                </table>
 +
</article>
 +
 
 +
<article id="item-2" class="menu__item">
 +
<table style="width:100%" class="calender">
 +
                    <tr>
 +
                        <th>Sun</th>
 +
                        <th>Mon</th>
 +
                        <th>Tue</th>
 +
                        <th>Wed</th>
 +
                        <th>Thu</th>
 +
                        <th>Fri</th>
 +
                        <th>Sat</th>
 +
                    </tr>
 +
                    <tr>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td class="td-item"><a href="#August-1">1</a></td>
 +
                        <td class="td-item"><a href="#August-2">2</a></td>
 +
                        <td class="td-item"><a href="#August-3">3</a></td>
 +
                        <td class="td-item"><a href="#August-4">4</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#August-5">5</a></td>
 +
                        <td class="td-item"><a href="#August-6">6</a></td>
 +
                        <td class="td-item"><a href="#August-7">7</a></td>
 +
                        <td class="td-item"><a href="#August-8">8</a></td>
 +
                        <td class="td-item"><a href="#August-9">9</a></td>
 +
                        <td class="td-item"><a href="#August-10">10</a></td>
 +
                        <td class="td-item"><a href="#August-11">11</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#August-12">12</a></td>
 +
                        <td class="td-item"><a href="#August-13">13</a></td>
 +
                        <td class="td-item"><a href="#August-14">14</a></td>
 +
                        <td class="td-item"><a href="#August-15">15</a></td>
 +
                        <td class="td-item"><a href="#August-16">16</a></td>
 +
                        <td class="td-item"><a href="#August-17">17</a></td>
 +
                        <td class="td-item"><a href="#August-18">18</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#August-19">19</a></td>
 +
                        <td class="td-item"><a href="#August-20">20</a></td>
 +
                        <td class="td-item"><a href="#August-21">21</a></td>
 +
                        <td class="td-item"><a href="#August-22">22</a></td>
 +
                        <td class="td-item"><a href="#August-23">23</a></td>
 +
                        <td class="td-item"><a href="#August-24">24</a></td>
 +
                        <td class="td-item"><a href="#August-25">25</a></td>
 +
                    </tr>
 +
                    <tr class="bottom">
 +
                        <td class="td-item"><a href="#August-26">26</a></td>
 +
                        <td class="td-item"><a href="#August-27">27</a></td>
 +
                        <td class="td-item"><a href="#August-28">28</a></td>
 +
                        <td class="td-item"><a href="#August-29">29</a></td>
 +
                        <td class="td-item"><a href="#August-30">30</a></td>
 +
                        <td class="td-item"><a href="#August-31">31</a></td>
 +
                        <td></td>
 +
                    </tr>
 +
                </table>
 +
</article>
 +
 
 +
<article id="item-3" class="menu__item">
 +
<table style="width:100%" class="calender">
 +
                    <tr>
 +
                        <th>Sun</th>
 +
                        <th>Mon</th>
 +
                        <th>Tue</th>
 +
                        <th>Wed</th>
 +
                        <th>Thu</th>
 +
                        <th>Fri</th>
 +
                        <th>Sat</th>
 +
                    </tr>
 +
                    <tr>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td class="td-item"><a href="#September-1">1</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#September-2">2</a></td>
 +
                        <td class="td-item"><a href="#September-3">3</a></td>
 +
                        <td class="td-item"><a href="#September-4">4</a></td>
 +
                        <td class="td-item"><a href="#September-5">5</a></td>
 +
                        <td class="td-item"><a href="#September-6">6</a></td>
 +
                        <td class="td-item"><a href="#September-7">7</a></td>
 +
                        <td class="td-item"><a href="#September-8">8</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#September-9">9</a></td>
 +
                        <td class="td-item"><a href="#September-10">10</a></td>
 +
                        <td class="td-item"><a href="#September-11">11</a></td>
 +
                        <td class="td-item"><a href="#September-12">12</a></td>
 +
                        <td class="td-item"><a href="#September-13">13</a></td>
 +
                        <td class="td-item"><a href="#September-14">14</a></td>
 +
                        <td class="td-item"><a href="#September-15">15</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#September-16">16</a></td>
 +
                        <td class="td-item"><a href="#September-17">17</a></td>
 +
                        <td class="td-item"><a href="#September-18">18</a></td>
 +
                        <td class="td-item"><a href="#September-19">19</a></td>
 +
                        <td class="td-item"><a href="#September-20">20</a></td>
 +
                        <td class="td-item"><a href="#September-21">21</a></td>
 +
                        <td class="td-item"><a href="#September-22">22</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#September-23">23</a></td>
 +
                        <td class="td-item"><a href="#September-24">24</a></td>
 +
                        <td class="td-item"><a href="#September-25">25</a></td>
 +
                        <td class="td-item"><a href="#September-26">26</a></td>
 +
                        <td class="td-item"><a href="#September-27">27</a></td>
 +
                        <td class="td-item"><a href="#September-28">28</a></td>
 +
                        <td class="td-item"><a href="#September-29">29</a></td>
 +
                    </tr>
 +
                    <tr class="bottom">
 +
                        <td class="td-item"><a href="#September-30">30</a></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                    </tr>
 +
                </table>
 +
</article>
 +
 
 +
<article id="item-4" class="menu__item">
 +
<table style="width:100%" class="calender">
 +
                    <tr>
 +
                        <th>Sun</th>
 +
                        <th>Mon</th>
 +
                        <th>Tue</th>
 +
                        <th>Wed</th>
 +
                        <th>Thu</th>
 +
                        <th>Fri</th>
 +
                        <th>Sat</th>
 +
                    </tr>
 +
                    <tr>
 +
                        <td></td>
 +
                        <td class="td-item"><a href="#October-1">1</a></td>
 +
                        <td class="td-item"><a href="#October-2">2</a></td>
 +
                        <td class="td-item"><a href="#October-3">3</a></td>
 +
                        <td class="td-item"><a href="#October-4">4</a></td>
 +
                        <td class="td-item"><a href="#October-5">5</a></td>
 +
                        <td class="td-item"><a href="#October-6">6</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#October-7">7</a></td>
 +
                        <td class="td-item"><a href="#October-8">8</a></td>
 +
                        <td class="td-item"><a href="#October-9">9</a></td>
 +
                        <td class="td-item"><a href="#October-10">10</a></td>
 +
                        <td class="td-item"><a href="#October-11">11</a></td>
 +
                        <td class="td-item"><a href="#October-12">12</a></td>
 +
                        <td class="td-item"><a href="#October-13">13</a></td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td class="td-item"><a href="#October-14">14</a></td>
 +
                        <td class="td-item"><a href="#October-15">15</a></td>
 +
                        <td class="td-item"><a href="#October-16">16</a></td>
 +
                        <td class="td-item"><a href="#October-17">17</a></td>
 +
                        <td class="td-item"><a href="#October-18">18</a></td>
 +
                        <td class="td-item"><a href="#October-19">19</a></td>
 +
                        <td>20</td>
 +
                    </tr>
 +
                    <tr>
 +
                        <td>21</td>
 +
                        <td>22</td>
 +
                        <td>23</td>
 +
                        <td>24</td>
 +
                        <td>25</td>
 +
                        <td>26</td>
 +
                        <td>27</td>
 +
                    </tr>
 +
                    <tr class="bottom">
 +
                        <td>28</td>
 +
                        <td>29</td>
 +
                        <td>30</td>
 +
                        <td>31</td>
 +
                        <td></td>
 +
                        <td></td>
 +
                        <td></td>
 +
                    </tr>
 +
                </table>
 +
</article>
 +
 
 +
</section>
 +
<div class="menu--shadow"></div>
 +
    </div>
 +
<div class="notebook">
 +
    <section class="notebook_wrapper">
 +
    <!---July------------------------------>
 +
    <article id="July-1" class="note-item note_active">
 +
        <h2>July 1</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="July-2" class="note-item">
 +
        <h2>July 2</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="July-3" class="note-item">
 +
        <h2>July 3</h2><hr>
 +
        <p class="note-title">none</p>
 +
    </article>
 +
    <article id="July-4" class="note-item">
 +
        <h2>July 4</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
 
 +
    <article id="July-5" class="note-item">
 +
        <h2>July 5</h2><hr>
 +
        <p class="note-title">Restore IDT in gblock fragment</p>
 +
        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
            <li class="list">Save DNA</li>
 +
            <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his-Sf1a NO.98351612</li>
 +
                      <li class="list">Sf1a-his- OAIP NO.98351613</li>
 +
                      <li class="list">OAIP-his- Hv1a NO.98351614</li>
 +
                      <li class="list">Hv1a-lectin-his NO.98351615</li>
 +
                      <li class="list">Sf1a-lectin-his NO.98351616</li>
 +
                      <li class="list">OAIP-lectin-his NO.98351617</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-6" class="note-item">
 +
        <h2>July 6</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="July-7" class="note-item">
 +
        <h2>July 7</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="July-8" class="note-item">
 +
        <h2>July 8</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
 
 +
    <article id="July-9" class="note-item">
 +
        <h2>July 9</h2><hr>
 +
        <p class="note-title">PCR of IDT gblock fragment</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The IDT PCR amplify the insert gene and use phusion DNA polymerase (PFU) to improve the PCR accuracy rate. </li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his-Sf1a</li>
 +
                      <li class="list">Sf1a-his- OAIP</li>
 +
                      <li class="list">OAIP-his- Hv1a</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check IDT PCR product</li>
 +
              <li class="list">Sample(7/9 PCR product)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-10" class="note-item">
 +
        <h2>July 10</h2><hr>
 +
        <p class="note-title">PCR of IDT gblock fragment</p>
 +
        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Amplify the DNA to revise the mistake.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his-Sf1a </li>
 +
                      <li class="list">Sf1a-his- OAIP</li>
 +
                      <li class="list">OAIP-his- Hv1a </li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check IDT PCR product</li>
 +
              <li class="list">Sample(7/10 PCR product)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
        <p class="note-title">Gel extraction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">For the purification of DNA, we cut the correct electrophoresis band to do gel extraction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the gene fragment(7/10) and pSB1C3 with EcoRI and PstI to ligase the gene fragment into the pSB1C3.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his 4 7.10 (EP)</li>
 +
                      <li class="list">Sf1a-his 1 7.10 (EP)</li>
 +
                      <li class="list">OAIP-his 2 7.10 (EP)</li>
 +
                      <li class="list">OAIP-lectin-his 5 7.10 (EP)</li>
 +
                      <li class="list">Hv1a-his 3 7.10 (EP) </li>
 +
                      <li class="list">Sf1a-lectin-his 6 7.10 (EP)</li>
 +
                      <li class="list">pSB1C3 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of digested product to check the pSB1C3 digestion.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">pSB1C3 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Ligation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Ligase DNA fragment into the pSB1C3</li>
 +
              <li class="list">Sample digestion product</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his 4 7.10 (EP)</li>
 +
                      <li class="list">Sf1a-his 1 7.10 (EP)</li>
 +
                      <li class="list">OAIP-his 2 7.10 (EP)</li>
 +
                      <li class="list">OAIP-lectin-his 5 7.10 (EP)</li>
 +
                      <li class="list">Hv1a-his 3 7.10 (EP)</li>
 +
                      <li class="list">Sf1a-lectin-his 6 7.10 (EP)</li>
 +
                      <li class="list">pSB1C3 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-11" class="note-item">
 +
        <h2>July 11</h2><hr>
 +
        <p class="note-title">Transformation:</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of ligated product to DH5α:</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 1 7.10 C<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his 4 7.10 C<sup>r</sup></li>
 +
                      <li class="list">OAIP-his 2 7.10 C<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his 5 7.10 C<sup>r</sup></li>
 +
                      <li class="list">Hv1a-his 3 7.10 C<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his 6 7.10 C<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-12" class="note-item">
 +
        <h2>July 12</h2><hr>
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of gel extraction product to check the concentration of gene to adjust the ligation protocol.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
        <p class="note-title">Transformation:</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of ligated product 20µL to BL21 competent cell 50µL</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 1 7.10 C<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his 4 7.10 C<sup>r</sup></li>
 +
                      <li class="list">OAIP-his 2 7.10 C<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his 5 7.10 C<sup>r</sup></li>
 +
                      <li class="list">Hv1a-his 3 7.10 C<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his 6 7.10 C<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-13" class="note-item">
 +
        <h2>July 13</h2><hr>
 +
        <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of ligated products to amplify the insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">OAIP-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">OAIP-lectin-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~8</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-14" class="note-item">
 +
        <h2>July 14</h2><hr>
 +
        <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of ligated products to amplify the insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-lectin-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~4</li>
 +
                </ul>
 +
        </ul>
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of PCR products to check the insert gene base pair</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-lectin-his C<sup>r</sup> 1~4</li>
 +
                </ul>
 +
        </ul>
 +
        <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of ligated product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his C<sup>r</sup> 3</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 4</li>
 +
                      <li class="list">OAIP-his C<sup>r</sup> 1</li>
 +
                      <li class="list">OAIP-lectin-his C<sup>r</sup> 3</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 3</li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 4</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-15" class="note-item">
 +
        <h2>July 15</h2><hr>
 +
        <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of ligated products to amplify the insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1~6</li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~3</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of PCR products to check the insert gene base pair</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~8</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1~6</li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1~3</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of Hv1a-lectin-his C<sup>r</sup>, Hv1a-his C<sup>r</sup> into BL21 again.</li>
 +
          </ul>
 +
 
 +
      <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have insert gene.(7/14)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his C<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his C<sup>r</sup></li>
 +
                      <li class="list">OAIP-his C<sup>r</sup></li>
 +
                    <li class="list">Hv1a-lectin-his C<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup></li>
 +
                    <li class="list">OAIP-lectin-his C<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-16" class="note-item">
 +
        <h2>July 16</h2><hr>
 +
        <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the plasmid (7/15) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 7.15 (EP)</li>
 +
                      <li class="list">OAIP-his 7.15 (EP)</li>
 +
                      <li class="list">Hv1a-his 7.15 (EP)</li>
 +
                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
 +
                      <li class="list">OAIP-lectin-his 7.15 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
              <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of digested product to check the band whether proper to the really base pair.</li>
 +
              <li class="list">Sample(7/16 Digestion product)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 7.15 (EP)</li>
 +
                      <li class="list">OAIP-his 7.15 (EP)</li>
 +
                      <li class="list">Hv1a-his 7.15 (EP)</li>
 +
                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
 +
                      <li class="list">OAIP-lectin-his 7.15 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of ligated products to amplify the insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1~11</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~7</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of PCR products to check the insert gene base pair.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1~11</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1~7</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of ligated product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 6</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 8</li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-17" class="note-item">
 +
        <h2>July 17</h2><hr>
 +
        <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the plasmid (7/15) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 7.15 (EP)</li>
 +
                      <li class="list">Hv1a-his 7.15 (EP)</li>
 +
                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
              <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of digested product to check the band whether proper to the really base pair.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 7.15 (EP)</li>
 +
                      <li class="list">Hv1a-his 7.15 (EP)</li>
 +
                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have insert gene.(7/16)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Sf1a-lectin-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 6</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 8</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
 
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of ligated product.(7/16)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 5</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 7</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 10</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 11</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 2</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 3</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 7</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
    <article id="July-18" class="note-item">
 +
        <h2>July 18</h2><hr>
 +
      <p class="note-title">PRC of IDT gblock fragment:</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Amplify the DNA to revise the mistake.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his-Sf1a NO.98351612</li>
 +
                      <li class="list">Sf1a-his-OAIP NO.98351613</li>
 +
                      <li class="list">OAIP-his-Hv1a NO.98351614</li>
 +
                      <li class="list">Hv1a-lectin-his NO.98351615</li>
 +
                      <li class="list">Sf1a-lectin-his NO.98351616</li>
 +
                      <li class="list">OAIP-lectin-his NO.98351617</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have insert gene.(7/17)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 5</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 7</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 10</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 11</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 3</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 7</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of ligated product.(7/17)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">OAIP-lectin-his C<sup>r</sup> 1</li>
 +
                      <li class="list">OAIP-lectin-his C<sup>r</sup> 2</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the plasmid (7/18) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his(EP)</li>
 +
                      <li class="list">Hv1a-his(EP)</li>
 +
                      <li class="list">Hv1a-his(EP)</li>
 +
                      <li class="list">Hv1a-his(EP)</li>
 +
                      <li class="list">Hv1a-lectin-his(EP)</li>
 +
                      <li class="list">Hv1a-lectin-his(EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Gel Electrophoresis of PCR-Amplified gBlocks</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list"></li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the gene fragment(7/18) and pSB1A3 with EcoRI and PstI to ligase the gene fragment into the pSB1A3.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 1 7.18 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 4 7.18 (EP)</li>
 +
                      <li class="list">OAIP-his 2 7.18 (EP)</li>
 +
                      <li class="list">OAIP-lectin-his 5 7.18 (EP)</li>
 +
                      <li class="list">Hv1a-his 3 7.18 (EP)</li>
 +
                      <li class="list">Sf1a-lectin-his 6 7.18 (EP)</li>
 +
                      <li class="list">pSB1C3 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of digested product to check the pSB1C3 digestion.</li>
 +
              <li class="list">Sample7/18 Digestion product</li>
 +
        </ul>
 +
 
 +
      <p class="note-title">Ligation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Ligase DNA fragment(7/18) into the pSB1C3</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
 
 +
                      <li class="list">Sf1a-his 1 7.18 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 4 7.18 (EP)</li>
 +
                      <li class="list">OAIP-his 2 7.18 (EP)</li>
 +
                      <li class="list">OAIP-lectin-his 5 7.18 (EP)</li>
 +
                      <li class="list">Hv1a-his 3 7.18 (EP)</li>
 +
                      <li class="list">Sf1a-lectin-his 6 7.18 (EP)</li>
 +
                      <li class="list">pSB1C3 (EP)</li>
 +
 
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">check the ligation product.</li>
 +
              <li class="list">Sample7/18 Ligation product</li>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-19" class="note-item">
 +
        <h2>July 19</h2><hr>
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of ligated product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of ligated product to DH5α.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 7/18 A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his 7/18 A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his 7/8 A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his 7/8 A<sup>r</sup></li>
 +
                      <li class="list">sf1a-lectin-his 7/8 A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his 7/18 A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have insert gene.(7/18)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">OAIP-lectin-his C<sup>r</sup> 7/19 1</li>
 +
                      <li class="list">OAIP-lectin-his C<sup>r</sup> 7/19 2</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the plasmid (7/19) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 2 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of ligated product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 7/19 1</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 7/19 2</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the plasmid (7/18) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 2</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 3</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 4</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 2</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of digestion product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 2</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 3</li>
 +
                      <li class="list">Hv1a-his C<sup>r</sup> 4</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 1</li>
 +
                      <li class="list">Hv1a-lectin-his C<sup>r</sup> 2</li>
 +
 
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-20" class="note-item">
 +
        <h2>July 20</h2><hr>
 +
      <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of ligated products(7/19) to amplify the insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1~4</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the gene fragment(7/18) and pSB1A3 with EcoRI and PstI to ligase the gene fragment into the pSB1A3.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 7/18 C<sup>r</sup>(EP)</li>
 +
                      <li class="list">Hv1a-his 7/18 C<sup>r</sup>(EP)</li>
 +
                      <li class="list">Hv1a-his 7/18 C<sup>r</sup>(EP)</li>
 +
                      <li class="list">Hv1a-his 7/18 C<sup>r</sup>(EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 7/18 C<sup>r</sup>(EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 7/18 C<sup>r</sup>(EP)</li>
 +
                      <li class="list">OAIP-lectin-his 7/18 C<sup>r</sup>(EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of digestion product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 1~4 7/18(EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 1, 2 7/18 (EP)</li>
 +
                      <li class="list">OAIP-lectin-his 7/19(EP)</li>
 +
                      <li class="list">Hv1a-his 3 7/18</li>
 +
                      <li class="list">Hv1a-lectin-his 2 7/18</li>
 +
                      <li class="list"></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check insert gene in E.coli.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 1-1~4</li>
 +
                      <li class="list">Hv1a-lectin-his 2-1~4</li>
 +
                      <li class="list">Sf1a-his3-1~4</li>
 +
                      <li class="list">Sf1a-lectin-his 4-1~4</li>
 +
                      <li class="list">OAIP-his 5-1~4</li>
 +
                      <li class="list">OAIP-lectin-his 6-1~4</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Sequencing</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">determine insert gene’s Sequence. </li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 3(The result is correct)</li>
 +
                      <li class="list">Hv1a-lectin-his 2</li>
 +
                      <li class="list">OAIP-lectin-his 1</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Ligation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Ligase DNA fragment(7/18) into the pSB1A3</li>
 +
              <li class="list">Sample digestion product</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 7/20 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 7/20 (EP)</li>
 +
                      <li class="list">OAIP-lectin-his 7/20 (EP)</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-21" class="note-item">
 +
        <h2>July 21</h2><hr>
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of ligated product.(7/20)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 2~4</li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have insert gene.(7/21)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 2~4</li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of ligated product(7/20) to <i>BL21-rosetta-gami</i></li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
 
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
    <article id="July-22" class="note-item">
 +
        <h2>July 22</h2><hr>
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the plasmid (7/21) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 2~4</li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of ligated product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 2~4</li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~4</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Sequencing</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">determine insert gene’s Sequence.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his (The result is correct)</li>
 +
                      <li class="list">Hv1a -his (The result is correct)</li>
 +
                      <li class="list">OAIP -his (The result is correct)</li>
 +
 
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the plasmid (7/21) with EcoRI and PstI to check whether the mini have insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 1~4(EP)</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4(EP)</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4(EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest IDT gBlock fragment for ES because lectin have P Cut position.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">Psb1A3</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of ligated product.</li>
 +
              <li class="list">Sample digestion product</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 1~4(EP)</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 1.3.4(EP)</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1.3.4(EP)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">PRC of IDT gblock fragment</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Amplify the DNA to revise the mistake.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Sf1a -his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
 
 +
 
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of digested product to check insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his (ES)</li>
 +
                      <li class="list">Sf1a-lectin-his (ES)</li>
 +
                      <li class="list">OAIP-lectin-his (ES)</li>
 +
                      <li class="list">[pSB1A3] (ES)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Ligation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Ligase DNA fragment(7/18) into the pSB1A3</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his (ES)</li>
 +
                      <li class="list">Sf1a-lectin-his (ES)</li>
 +
                      <li class="list">OAIP-lectin-his (ES)</li>
 +
                      <li class="list">[pSB1A3] (ES)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of backbone plasmid into <i>BL21-rosetta-gami</i> check whether the competent cell is working.</li>
 +
              <li class="list">we choose pSB1A3 to transformation because <i>BL21-rosetta-gami</i> has C<sup>r</sup> and K<sup>r</sup>.</li>
 +
 
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-23" class="note-item">
 +
        <h2>July 23</h2><hr>
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">After the PRC of IDT gBlock fragment, we electrophoresis samples to check the insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">There is a fragment in the snow drop lectin identification with PstI. Therefore, if sequence have snow drop lectin, we digested the gene fragment(7/22) and pSB1A3 with EcoRI and Spe l to ligase the gene fragment into the pSB1A3. Others still digested with EcoRI and PstI.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 3 7/18 (EP)</li>
 +
                      <li class="list">Sf1a-his 1 7/8 (EP)</li>
 +
                      <li class="list">OAIP-his 2 7/18 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 4 7/8 (ES)</li>
 +
                      <li class="list">Sf1a-lectin-his 6 7/18 (ES)</li>
 +
                      <li class="list">OAIP-lectin-his 5 7/18(ES)</li>
 +
                      <li class="list">pSB1A3 (EP) </li>
 +
                      <li class="list">pSB1A3 (ES) </li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Ligation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Ligase DNA fragment(7/23) into the pSB1A3</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 3 7/18 (EP)</li>
 +
                      <li class="list">Sf1a-his 1 7/8 (EP)</li>
 +
                      <li class="list">OAIP-his 2 7/18 (EP)</li>
 +
                      <li class="list">Hv1a-lectin-his 4 7/8 (ES)</li>
 +
                      <li class="list">Sf1a-lectin-his 6 7/18 (ES)</li>
 +
                      <li class="list">OAIP-lectin-his 5 7/18(ES)</li>
 +
                      <li class="list">pSB1A3 (EP) </li>
 +
                      <li class="list">pSB1A3 (ES) </li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of ligated product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/22</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/22</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/22</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-24" class="note-item">
 +
        <h2>July 24</h2><hr>
 +
      <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of ligated products(7/23) to amplify the insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/23 1~4</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/23 5~8</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/23 9~12</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of PCR products</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/3 1~4</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/23 5~8</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/23 9~12</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of ligated product.(7/24)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/24 2.3</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/24 5.7</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/24 9.12</li>
 +
 
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-25" class="note-item">
 +
        <h2>July 25</h2><hr>
 +
      <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have insert gene.(7/24)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 7/24 2.3</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 7/24 5.7</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 7/24 9.12</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest the plasmid (7/25) with EcoRI and Spel to check whether the mini have insert gene fragment.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup>  2.3(ES)</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup>  5.7(ES)</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup>  9.12(ES)</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
        <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of ligated product.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup>  2.3(ES)</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup>  5.7(ES)</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup>  9.12(ES)</li>
 +
 
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Sequencing</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">determine insert gene’s Sequence.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his (The result is correct)</li>
 +
                      <li class="list">Hv1a-lectin-his (The result is correct)</li>
 +
                      <li class="list">OAIP - lectin-his (The result is correct)</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-26" class="note-item">
 +
        <h2>July 26</h2><hr>
 +
      <p class="note-title">Prepare of CaCl<sub>2</sub></p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">for competent cell</li>
 +
          </ul>
 +
 
 +
        <p class="note-title">Rosetta-gami competent cell prepare</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">for expression of disulfide bond.</li>
 +
          </ul>
 +
 
 +
        <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of backbone plasmid into <i>BL21-rosetta-gami</i> check whether the competent cell is working.</li>
 +
              <li class="list">we choose pSB1A3 to transformation because <i>BL21-rosetta-gami</i> has C<sup>r</sup> and K<sup>r</sup>.</li>
 +
              <li class="list">Only write colonies. NO RFP EXPRESSION</li>
 +
          </ul>
 +
    </article>
 +
 
 +
    <article id="July-27" class="note-item">
 +
        <h2>July 27</h2><hr>
 +
        <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of backbone plasmid into <i>BL21-rosetta-gami</i> check whether the competent cell is working.</li>
 +
              <li class="list">we choose pSB1A3 to transformation because <i>BL21-rosetta-gami</i> has C<sup>r</sup> and K<sup>r</sup>.</li>
 +
              <li class="list">Only write colonies. NO RFP EXPRESSION</li>
 +
          </ul>
 +
 
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of plasmid which have correct sequence to DH5α.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a -his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">Hv1a -his</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-28" class="note-item">
 +
        <h2>July 28</h2><hr>
 +
        <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of correct colonies. (7/27)</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Sf1a -his A<sup>r</sup></li>
 +
                        <li class="list">OAIP-his A<sup>r</sup></li>
 +
                        <li class="list">Hv1a -his A<sup>r</sup></li>
 +
                    </ul>
 +
          </ul>
 +
 
 +
        <p class="note-title">Test competent cell</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list"><i>BL21-rosetta-gami</i> aren’t express RFP but write colonies. Therefore, we test competent cell whether they could save on A<sup>r</sup> agar plate.</li>
 +
              <li class="list">After cultivation about 14 hours, there are many write colonies. We think them may be contamination during preparing competent cell.</li>
 +
          </ul>
 +
    </article>
 +
 
 +
    <article id="July-29" class="note-item">
 +
        <h2>July 29</h2><hr>
 +
      <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have correct insert gene(7/28) to conserve and transform to <i>BL21-Rosetta-gami</i>.</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Sf1a -his A<sup>r</sup></li>
 +
                        <li class="list">OAIP-his A<sup>r</sup></li>
 +
                        <li class="list">Hv1a -his A<sup>r</sup></li>
 +
                    </ul>
 +
          </ul>
 +
 
 +
 
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of plasmid which have correct sequence to DH5α.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his 7/24 A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his 7/24 A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his 7/24 A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="July-30" class="note-item">
 +
        <h2>July 30</h2><hr>
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of correct colonies. (7/29)</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Sf1a-lectin-his 7/29 A<sup>r</sup></li>
 +
                        <li class="list">OAIP-lectin-his 7/29 A<sup>r</sup></li>
 +
                        <li class="list">Hv1a-lectin-his 7/29 A<sup>r</sup></li>
 +
                    </ul>
 +
          </ul>
 +
 
 +
 
 +
      <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have correct insert gene(7/30) to conserve and transform to  BL21-Rosetta-gami.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Sf1a-lectin-his 7/29 A<sup>r</sup></li>
 +
                        <li class="list">OAIP-lectin-his 7/29 A<sup>r</sup></li>
 +
                        <li class="list">Hv1a-lectin-his 7/29 A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Preparation of Competent cell</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Rosetta-gami competent cell:</li>
 +
              <li class="list">Prepare the bacteria that can make disulfide bond</li>
 +
 
 +
        </ul>
 +
    </article>
 +
    <article id="July-31" class="note-item">
 +
        <h2>July 31</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
 
 +
    <!-------------------------------------August------------------------------------------------->
 +
 
 +
    <article id="August-1" class="note-item">
 +
        <h2>August 1</h2><hr>
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of pSB1A3 to test the competent cell work.</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">PSB1A3</li>
 +
                    </ul>
 +
          </ul>
 +
 
 +
 
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Hv1a-his</li>
 +
                        <li class="list">Sf1a-his</li>
 +
                        <li class="list">OAIP-his</li>
 +
                        <li class="list">Hv1a-lectin-his</li>
 +
                        <li class="list">Sf1a-lectin-his</li>
 +
                        <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-2" class="note-item">
 +
        <h2>August 2</h2><hr>
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Sf1a-his 7/28 A<sup>r</sup></li>
 +
                        <li class="list">OAIP-his 7/28 A<sup>r</sup></li>
 +
                        <li class="list">Hv1a-lectin-his 7/29 A<sup>r</sup></li>
 +
                        <li class="list">Sf1a-lectin-his 7/29 A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-3" class="note-item">
 +
        <h2>August 3</h2><hr>
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of <i>BL21-Rosetta-gam</i> colonies for IPTG induction.</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Hv1a-his 8/1 A<sup>r</sup></li>
 +
                        <li class="list">OAIP-lectin-his 8/1 A<sup>r</sup></li>
 +
                    </ul>
 +
          </ul>
 +
 
 +
 
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of correct colonies again for mini extraction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Sf1a-his 7/27 A<sup>r</sup></li>
 +
                        <li class="list">OAIP-his 7/27 A<sup>r</sup></li>
 +
                        <li class="list">Hv1a-lectin-his 7/24 A<sup>r</sup></li>
 +
                        <li class="list">Sf1a-lectin-his 7/24 A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-4" class="note-item">
 +
        <h2>August 4</h2><hr>
 +
      <p class="note-title">Miniprep</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purify plasmid which have correct insert gene(8/3) to conserve and transform to <i>BL21-Rosetta-gami</i>.</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">OAIP-his 8/3  A<sup>r</sup></li>
 +
                        <li class="list">Sf1a-lectin-his 8/3 A<sup>r</sup></li>
 +
                        <li class="list">Hv1a-lectin-his 8/3 A<sup>r</sup></li>
 +
                    </ul>
 +
          </ul>
 +
 
 +
 
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of correct colonies again for mini extraction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Sf1a-his 7/27 A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
 
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">OAIP-his 8/3 A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his 8/3 A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his 8/3 A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">To find out the best condition for IPTG induction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 8/3 A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his 8/3 A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Condition<br>0μM, 250μM, 500μM, 750μM, 1000μM</li>
 +
              <li class="list">Add IPTG concentration and test OD<sub>600</sub><br>0μM, 250μM, 500μM, 750μM, 1000μM</li>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-5" class="note-item">
 +
        <h2>August 5</h2><hr>
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the protein production after IPTG condition.</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Marker</li>
 +
                        <li class="list">OAIP-lectin-his 250μM 2hr</li>
 +
                        <li class="list">OAIP-lectin-his 500μM 2hr</li>
 +
                        <li class="list">OAIP-lectin-his 750μM 2hr</li>
 +
                        <li class="list">OAIP-lectin-his 1000μM 2hr</li>
 +
                        <li class="list">OAIP-lectin-his 0μM 4hr</li>
 +
                        <li class="list">OAIP-lectin-his 250μM 4hr</li>
 +
                        <li class="list">OAIP-lectin-his 500μM 4hr</li>
 +
                        <li class="list">OAIP-lectin-his 750μM 4hr</li>
 +
                        <li class="list">OAIP-lectin-his 1000μM 4hr</li>
 +
                        <li class="list">Marker</li>
 +
                        <li class="list">OAIP-lectin-his 250μM 6hr</li>
 +
                        <li class="list">OAIP-lectin-his 500μM 6hr</li>
 +
                        <li class="list">OAIP-lectin-his 750μM 6hr</li>
 +
                        <li class="list">OAIP-lectin-his 1000μM 6hr</li>
 +
                        <li class="list">OAIP-lectin-his 0μM 8hr</li>
 +
                        <li class="list">OAIP-lectin-his 250μM 8hr</li>
 +
                        <li class="list">OAIP-lectin-his 500μM 8hr</li>
 +
                        <li class="list">OAIP-lectin-his 750μM 8hr</li>
 +
                        <li class="list">OAIP-lectin-his 1000μM 8hr</li>
 +
                    </ul>
 +
          </ul>
 +
    </article>
 +
 
 +
 
 +
    <article id="August-6" class="note-item">
 +
        <h2>August 6</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="August-7" class="note-item">
 +
        <h2>August 7</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="August-8" class="note-item">
 +
        <h2>August 8</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="August-9" class="note-item">
 +
        <h2>August 9</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="August-10" class="note-item">
 +
        <h2>August 10</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="August-11" class="note-item">
 +
        <h2>August 11</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="August-12" class="note-item">
 +
        <h2>August 12</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="August-13" class="note-item">
 +
        <h2>August 13</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
 
 +
 
 +
    <article id="August-14" class="note-item">
 +
        <h2>August 14</h2><hr>
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 7/29  A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his 8/4 A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-his 7/29  A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his 8/4  A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his 7/30  A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his 8/4  A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of transformation colonies to check the base pair.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 1~6</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 1~6</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 1~6</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 1~6</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-15" class="note-item">
 +
        <h2>August 15</h2><hr>
 +
      <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of ligated products to amplify the insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 8/14  A<sup>r</sup> 1~6</li>
 +
                      <li class="list">Hv1a-lectin-his 8/14 A<sup>r</sup> 1~6</li>
 +
                      <li class="list">Sf1a- lectin-his 8/14  A<sup>r</sup> 1~6</li>
 +
                      <li class="list">Sf1a-lectin-his 8/4  A<sup>r</sup> 1~6</li>
 +
                      <li class="list">OAIP-lectin-his 7/30  A<sup>r</sup> 1~6</li>
 +
                      <li class="list">OAIP- lectin-his 8/14  A<sup>r</sup> 1~6</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of PCR products to check the insert gene base pair</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 8/15 A<sup>r</sup> 1~6</li>
 +
                      <li class="list">Hv1a-lectin-his 8/15 A<sup>r</sup> 1~6</li>
 +
                      <li class="list">Sf1a- lectin-his 8/15 A<sup>r</sup> 1~6</li>
 +
                      <li class="list">OAIP- lectin-his 8/15 A<sup>r</sup> 1~6</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of plasmid which have correct sequence to <i>BL21-Rosetta-gami</i>.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his 7/29 A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his 8/4 A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">PCR of ligated products to amplify the insert gene.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a- lectin-his 8/15 A<sup>r</sup> 1~6</li>
 +
                      <li class="list">OAIP- lectin-his 8/15 A<sup>r</sup> 1~6</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of PCR products to check the insert gene base pair.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a- lectin-his 8/15 A<sup>r</sup> 1~6</li>
 +
                      <li class="list">OAIP- lectin-his 8/15 A<sup>r</sup> 1~6</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the production of IPTG induction (8/4) again.</li>
 +
              <li class="list">No target protein expression.</li>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-16" class="note-item">
 +
        <h2>August 16</h2><hr>
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of <i>BL21-Rosetta-gami</i> colonies for IPTG induction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 8/15  A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his 8/15 A<sup>r</sup></li>
 +
                      <li class="list">Sf1a- lectin-his 8/15  A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his 8/15  A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">To check the protein expression for IPTG induction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 8/16  A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his 8/16 A<sup>r</sup></li>
 +
                      <li class="list">Sf1a- lectin-his 8/16  A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his 8/16 A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Condition<br>0μM, 500μM</li>
 +
              <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
 +
        </ul>
 +
 
 +
      <p class="note-title">Sonication and SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.</li>
 +
              <li class="list">Sample(8/16)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 0μM 2hr</li>
 +
                      <li class="list">Hv1a-his 500μM 2hr</li>
 +
                      <li class="list">Sf1a-lectin-his 0μM 4hr</li>
 +
                      <li class="list">Sf1a-lectin-his 500μM 4hr</li>
 +
                      <li class="list">Hv1a-lectin-his 0μM 2hr</li>
 +
                      <li class="list">Hv1a-lectin-his 500μM 2hr</li>
 +
                      <li class="list">OAIP-lectin-his 0μM 4hr</li>
 +
                      <li class="list">OAIP-lectin-his 500μM 4hr</li>
 +
                </ul>
 +
              <li class="list">Failed. Do it again</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 0μM 2hr</li>
 +
                      <li class="list">Hv1a-his 500μM 2hr</li>
 +
                      <li class="list">Sf1a-lectin-his 0μM 4hr</li>
 +
                      <li class="list">Sf1a-lectin-his 500μM 4hr</li>
 +
                      <li class="list">Hv1a-lectin-his 0μM 2hr</li>
 +
                      <li class="list">Hv1a-lectin-his 500μM 2hr</li>
 +
                      <li class="list">OAIP-lectin-his 0μM 4hr</li>
 +
                      <li class="list">OAIP-lectin-his 500μM 4hr</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-17" class="note-item">
 +
        <h2>August 17</h2><hr>
 +
          <p class="note-title">IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">To check the protein expression for IPTG induction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 8/17 A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his </li>
 +
                      <li class="list">Sf1a- lectin-his</li>
 +
                      <li class="list">OAIP- lectin-his</li>
 +
 
 +
                </ul>
 +
              <li class="list">Condition<br>
 +
0μM, 500μM
 +
</li>
 +
              <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
 +
        </ul>
 +
 
 +
 
 +
      <p class="note-title">Sonication and SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 0μM 2hr</li>
 +
                      <li class="list">Hv1a-his 500μM 2hr</li>
 +
                      <li class="list">Sf1a-lectin-his 0μM 4hr</li>
 +
                      <li class="list">Sf1a-lectin-his 500μM 4hr</li>
 +
                      <li class="list">Hv1a-lectin-his 0μM 2hr</li>
 +
                      <li class="list">Hv1a-lectin-his 500μM 2hr</li>
 +
                      <li class="list">OAIP-lectin-his 0μM 4hr</li>
 +
                      <li class="list">OAIP-lectin-his 500μM 4hr</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-18" class="note-item">
 +
        <h2>August 18</h2><hr>
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">To Check protein disulfide bond folding.</li>
 +
              <li class="list">β-me make disulfide bond not folding.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his 0μM + β-me</li>
 +
                      <li class="list">Hv1a-lectin-his 500μM + β-me</li>
 +
                      <li class="list">Hv1a-lectin-his 500μM</li>
 +
                      <li class="list">Sf1a-lectin-his 0μM + β-me</li>
 +
                      <li class="list">Sf1a-lectin-his 500μM + β-me</li>
 +
                      <li class="list">Sf1a-lectin-his 500μM</li>
 +
                      <li class="list">OAIP-lectin-his 0μM + β-me</li>
 +
                      <li class="list">OAIP-lectin-his 500μM + β-me</li>
 +
                      <li class="list">OAIP-lectin-his 500μM</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-19" class="note-item">
 +
        <h2>August 19</h2><hr>
 +
        <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">To Check protein disulfide bond folding with the induction of control, Rosetta-gami stain.</li>
 +
              <li class="list">β-me make disulfide bond not folding.</li>
 +
              <li class="list">Sample(8/17)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his 0μM + β-me</li>
 +
                      <li class="list">Hv1a-lectin-his 500μM + β-me</li>
 +
                      <li class="list">Hv1a-lectin-his 500μM</li>
 +
                      <li class="list">Sf1a-lectin-his 0μM + β-me</li>
 +
                      <li class="list">Sf1a-lectin-his 500μM + β-me</li>
 +
                      <li class="list">Sf1a-lectin-his 500μM</li>
 +
                      <li class="list">Rosetta-gami + β-me</li>
 +
                      <li class="list">OAIP-lectin-his 0μM + β-me</li>
 +
                      <li class="list">OAIP-lectin-his 500μM + β-me</li>
 +
                      <li class="list">OAIP-lectin-his 500μM</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 
 +
    <article id="August-20" class="note-item">
 +
        <h2>August 20</h2><hr>
 +
      <p class="note-title">Sonication and SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/20</li>
 +
                      <li class="list">Hv1a-lectin-his  A<sup>r</sup> 8/20</li>
 +
                      <li class="list">Sf1a-lectin-hiss A<sup>r</sup> 8/20</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/20</li>
 +
                </ul>
 +
        </ul>
 +
 
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">To Check protein disulfide bond folding with the induction of control, <i>Rosetta-gami stain</i>.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his 8/19</li>
 +
                      <li class="list">Sf1a-his 8/19</li>
 +
                      <li class="list">OAIP-his 8/19</li>
 +
                </ul>
 
</ul>
 
</ul>
 +
    </article>
 +
 +
    <article id="August-21" class="note-item">
 +
        <h2>August 21</h2><hr>
 +
      <p class="note-title">IPTG Induction and Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">To check the protein expression for IPTG induction. (Second round)</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/20</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/20</li>
 +
                      <li class="list">Sf1a-his  A<sup>r</sup> 8/17</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/20</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 8/17</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/20</li>
 +
                </ul>
 +
              <li class="list">Condition<br>0μM, 500μM</li>
 +
              <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-22" class="note-item">
 +
        <h2>August 22</h2><hr>
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list"> Hv1a-lectin-his 8/20  A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list"><i>BL21-Rosetta-gami</i> competent cell</li>
 +
                      <li class="list">Cultivation 11 hours</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">IPTG Induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Prepare for the purify the protein from sample <i>BL21-Rosetta-gami</i> colonies.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his Ar 8/20 A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Condition<br>0μM, 1000μM</li>
 +
              <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his 8/22</li>
 +
                      <li class="list">BL21-Rosetta-gami 8/22</li>
 +
                      <li class="list">Sf1a-lectin-his 0μM 8/22</li>
 +
                      <li class="list">Sf1a-lectin-his 1000μM 8/22</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-23" class="note-item">
 +
        <h2>August 23</h2><hr>
 +
      <p class="note-title">Protein purification(8/22-8/23)</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation and IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/20</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/20</li>
 +
                </ul>
 +
              <li class="list">Add 1000μM IPTG and test OD<sub>600</sub> </li>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/23</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/23</li>
 +
                </ul>
 +
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-24" class="note-item">
 +
        <h2>August 24</h2><hr>
 +
      <p class="note-title">Protein purification(8/23-8/24)</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation and IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 8/20</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/20</li>
 +
                </ul>
 +
              <li class="list">Refresh to approximately OD<sub>600</sub> 0.7</li>
 +
              <li class="list">Add 1000μM IPTG and test OD<sub>600</sub></li>
 +
              <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his: 0.966</li>
 +
                      <li class="list">Sf1a-lectin-his: 1.363</li>
 +
                </ul>
 +
              <li class="list">Certification, and add 2mL binding buffer and 200μM PMSF for sonication. Repeat three times.</li>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 8/24</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/24</li>
 +
                </ul>
 +
 +
        </ul>
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample</li>
 +
              <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his(8/22-8/23)</li>
 +
              </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-25" class="note-item">
 +
        <h2>August 25</h2><hr>
 +
      <p class="note-title">Protein purification(8/24-8/25)</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample</li>
 +
              <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his(8/23-8/24)</li>
 +
              </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-26" class="note-item">
 +
        <h2>August 26</h2><hr>
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/15</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/15</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/15</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/15</li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 8/24</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/24</li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 8/17</li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> 8/17</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Induce the mass protein expression for SDS-PAGE.</li>
 +
              <li class="list">The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/26</li>
 +
                </ul>
 +
              <li class="list">Add 500μM IPTG</li>
 +
              <li class="list">Take the following samples to go on the experience first. The others storage in 4℃ refrigerator.</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the protein in pellet or in solution.</li>
 +
              <li class="list">Sample</li>
 +
              <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/26(with urea)</li>
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26(with urea)</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
 +
              </ul>
 +
              <li class="list">Sonication and the supernatant is loading sample1</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">10 seconds on, 40 seconds off and repeat six times.</li>
 +
                </ul>
 +
              <li class="list">Add binding buffer (with/without urea) again and the solution is loading sample 2</li>
 +
              <li class="list">Sonication again and the supernatant is loading sample 3</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">10 seconds on, 40 seconds off and repeat six times.</li>
 +
                </ul>
 +
              <li class="list">Add binding buffer (with/without urea) again and the solution is loading sample4</li>
 +
              <li class="list">Loading sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his (with urea) 1~4</li>
 +
                      <li class="list">Hv1a-his 1~4</li>
 +
                      <li class="list">Hv1a-lectin-his (with urea) 1~4</li>
 +
                      <li class="list">Hv1a-lectin-his 1~4</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Induce the mass protein expression for SDS-PAGE.</li>
 +
              <li class="list">The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/26</li>
 +
                </ul>
 +
              <li class="list">Add 500μM IPTG and test OD<sub>600</sub> to approximately 1.050</li>
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the protein in pellet or in solution.</li>
 +
              <li class="list">Sample</li>
 +
              <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/26(with urea)</li>
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26(with urea)</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/26(with urea)</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/26</li>
 +
                      <li class="list">OAIPlectin-his A<sup>r</sup> 8/26(with urea)</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/26</li>
 +
              </ul>
 +
              <li class="list">Certificate the sample, add binding buffer (with/without urea), PMSF</li>
 +
              <li class="list">Sonication and the supernatant is loading sample1</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">10 seconds on, 40 seconds off and repeat six times.</li>
 +
                </ul>
 +
              <li class="list">Add binding buffer (with/without urea) again and the solution is loading sample 2</li>
 +
              <li class="list">Sonication again and the supernatant is loading sample 3</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">10 seconds on, 40 seconds off and repeat six times.</li>
 +
                </ul>
 +
              <li class="list">Add binding buffer (with/without urea) again and the solution is loading sample4</li>
 +
              <li class="list">Loading sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his (with urea) 1~4</li>
 +
                      <li class="list">Hv1a-his 1~4</li>
 +
                      <li class="list">Hv1a-lectin-his (with urea) 1~4</li>
 +
                      <li class="list">Hv1a-lectin-his 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his (with urea) 1~4</li>
 +
                      <li class="list">Sf1a-lectin-his 1~4</li>
 +
                      <li class="list">OAIPlectin-his (with urea) 1~4</li>
 +
                      <li class="list">OAIP-lectin-his 1~4</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-27" class="note-item">
 +
        <h2>August 27</h2><hr>
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> </li>
 +
                      <li class="list">Hv1a-lectin-his  A<sup>r</sup> </li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
    <article id="August-28" class="note-item">
 +
        <h2>August 28</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample(8/27)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample(8/28)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 8/17</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-29" class="note-item">
 +
        <h2>August 29</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample(8/28)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample(8/29)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup> 8/17</li>
 +
                </ul>
 +
        </ul>
 +
 +
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> 8/15</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/15</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/15</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/15</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-30" class="note-item">
 +
        <h2>August 30</h2><hr>
 +
      <p class="note-title">IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Induce the mass protein expression for SDS-PAGE.</li>
 +
              <li class="list">The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.</li>
 +
              <li class="list">Sample</li>
 +
              <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/29</li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/29</li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup> 8/29</li>
 +
 +
              </ul>
 +
              <li class="list">Refresh to approximately OD<sub>600</sub> 0.6</li>
 +
              <li class="list">Add 500μM IPTG and cultivate for 4hr</li>
 +
        </ul>
 +
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample(8/30)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample(8/29)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
      <p class="note-title">PCR</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">amplify the insert gene</li>
 +
              <li class="list">Sample(8/30)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of PCR product</li>
 +
              <li class="list">Sample(8/30 PCR product)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Digestion</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Digest min for transformation</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his digestion EP</li>
 +
                      <li class="list">Hv1a-lectin-his digestion ES</li>
 +
                      <li class="list">Sf1a-his digestion EP</li>
 +
                      <li class="list">Sf1a-lectin-his digestion ES</li>
 +
                      <li class="list">OAIP-his digestion EP</li>
 +
                      <li class="list">OAIP-lectin-his digestion ES</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation PSB1C3 to DH5α</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Trans PSB1C3 8/30</li>
 +
                      <li class="list">Trans PSB1C3 8/30</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Urea test</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check whether protein in inclusion body or not.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-his + Urea</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-his + Urea</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-his + Urea</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="August-31" class="note-item">
 +
        <h2>August 31</h2><hr>
 +
      <p class="note-title">Ligation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Ligase insert gene into the pSB1C3</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Transformation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Transformation of ligated product to DH5α</li>
 +
              <li class="list">Sample(8/31 Ligation product)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Dialysis and refolding</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Dialysis and refolding for insoluble protein</li>
 +
              <li class="list">Sample(8/8)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his (8/30)</li>
 +
                      <li class="list">Sf1a-lectin-his(8/30)</li>
 +
                      <li class="list">OAIP-lectin-his(8/30)</li>
 +
                      <li class="list">Hv1a-his(8/28)</li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample(8/30)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
 +
        </ul>
 +
    </article>
 +
 +
    <!--------------------------------September----------------------------------->
 +
 +
    <article id="September-1" class="note-item">
 +
        <h2>September 1</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample(8/30)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                      <li class="list">Hv1a-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample(8/30)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                      <li class="list">Hv1a-his</li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> </li>
 +
                      <li class="list">Hv1a-lectin-his  A<sup>r</sup> </li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> </li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup> </li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">IPTG Induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Prepare for the purify the protein from sample <i>BL21-Rosetta-gami</i> colonies.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> </li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> </li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup> </li>
 +
                </ul>
 +
              <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup> </li>
 +
                      <li class="list">OAIP-his A<sup>r</sup> </li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup> </li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample <i>BL21-Rosetta-gami</i> colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-2" class="note-item">
 +
        <h2>September 2</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Dialysis and refolding</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Dialysis and refolding for insoluble protein</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-3" class="note-item">
 +
        <h2>September 3</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
                <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
      <p class="note-title">Concentrate protein</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Increase the concentration of protein</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-4" class="note-item">
 +
        <h2>September 4</h2><hr>
 +
        <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-5" class="note-item">
 +
        <h2>September 5</h2><hr>
 +
      <p class="note-title">IPTG Induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Prepare for the purify the protein from sample <i>BL21-Rosetta-gami</i> colonies.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Add IPTG 500μM and test OD<sub>600</sub> after 4hr.</li>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">UV test</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Test degradation rate</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Concentrate protein</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Increase the concentration of protein</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-6" class="note-item">
 +
        <h2>September 6</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-7" class="note-item">
 +
        <h2>September 7</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
 +
      <p class="note-title">UV test</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Test degradation rate</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-8" class="note-item">
 +
        <h2>September 8</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
 +
 +
      <p class="note-title">Cultivation and IPTG induction</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Cultivation of sample BL21-Rosetta-gami colonies for protein purification.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Refresh to approximately OD<sub>600</sub> 0.7</li>
 +
              <li class="list">Add 1000μM IPTG and test OD<sub>600</sub> </li>
 +
                  <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his:0.772</li>
 +
                      <li class="list">Hv1a-lectin-his:0.631</li>
 +
                      <li class="list">Sf1a-his:0.567</li>
 +
                      <li class="list">Sf1a-lectin-his:0.78</li>
 +
                      <li class="list">OAIP-his:0.851</li>
 +
                      <li class="list">OAIP-lectin-his:0.819</li>
 +
                </ul>
 +
              <li class="list">Certification, and add 2mL binding buffer and 200μM PMSF for sonication. Repeat three times.</li>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Sonication</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-9" class="note-item">
 +
        <h2>September 9</h2><hr>
 +
      <p class="note-title">Protein purification</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">The buffer contains the urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
              <li class="list">Collect：<br>pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH<sub>2</sub>O solution.</li>
 +
        </ul>
 +
      <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
 +
        </ul>
 +
    </article>
 +
 +
 +
 +
<!--9/10-->
 +
    <article id="September-10" class="note-item">
 +
        <h2>September 10</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-11" class="note-item">
 +
        <h2>September 11</h2><hr>
 +
      <p class="note-title">Dialysis protein</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Dialysis and refolding for insoluble protein</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his  E1-E3</li>
 +
                      <li class="list">Hv1a-lectin-his  E1-E5</li>
 +
                      <li class="list">Sf1a-his  E1-E5</li>
 +
                      <li class="list">Sf1a-lectin-his E1-E4</li>
 +
                      <li class="list">OAIP-his  E1-E3</li>
 +
                      <li class="list">OAIP-lectin-his  E1-E2</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-12" class="note-item">
 +
        <h2>September 12</h2><hr>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-13" class="note-item">
 +
        <h2>September 13</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-14" class="note-item">
 +
        <h2>September 14</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-15" class="note-item">
 +
        <h2>September 15</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-16" class="note-item">
 +
        <h2>September 16</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-17" class="note-item">
 +
        <h2>September 17</h2><hr>
 +
      <p class="note-title">Dialysis protein</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Dialysis and refolding for insoluble protein</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his  E2-E3</li>
 +
                      <li class="list">Hv1a-lectin-his  E1-E6</li>
 +
                      <li class="list">Sf1a-his  E1-E4</li>
 +
                      <li class="list">Sf1a-lectin-his E2-E4</li>
 +
                      <li class="list">OAIP-his  E2-E4</li>
 +
                      <li class="list">OAIP-lectin-his  E2-E3</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-18" class="note-item">
 +
        <h2>September 18</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-19" class="note-item">
 +
        <h2>September 19</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-20" class="note-item">
 +
        <h2>September 20</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-21" class="note-item">
 +
        <h2>September 21</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-22" class="note-item">
 +
        <h2>September 22</h2><hr>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-23" class="note-item">
 +
        <h2>September 23</h2><hr>
 +
      <p class="note-title">Dialysis protein</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Remove urea</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his E1-E3</li>
 +
                      <li class="list">Hv1a-lectin-his E1-E6</li>
 +
                      <li class="list">Sf1a-his E1-E4</li>
 +
                      <li class="list">Sf1a-lectin-his E1-E6</li>
 +
                      <li class="list">OAIP-his E1-E3</li>
 +
                      <li class="list">OAIP-lectin-his E1-E6</li>
 +
                </ul>
 +
        </ul>
 +
 +
              <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein again</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his </li>
 +
                      <li class="list">Hv1a-lectin-his (no β-me)</li>
 +
                      <li class="list">Sf1a-his </li>
 +
                      <li class="list">Sf1a-lectin-his (no β-me)</li>
 +
                      <li class="list">OAIP-his </li>
 +
                      <li class="list">OAIP-lectin-his (no β-me)</li>
 +
                </ul>
 +
 +
        </ul>
 +
 +
 +
              <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of composite parts again</li>
 +
              <li class="list">Sample(8/30 PCR product)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-24" class="note-item">
 +
        <h2>September 24</h2><hr>
 +
              <p class="note-title">Electrophoresis</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Electrophoresis of composite parts again</li>
 +
              <li class="list">Sample(8/30 PCR product)</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-25" class="note-item">
 +
        <h2>September 25</h2><hr>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-lectin-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="September-26" class="note-item">
 +
        <h2>September 26</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-27" class="note-item">
 +
        <h2>September 27</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-28" class="note-item">
 +
        <h2>September 28</h2><hr>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his A<sup>r</sup></li>
 +
                      <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
 +
                      <li class="list">Sf1a-his A<sup>r</sup></li>
 +
                      <li class="list">OAIP-his A<sup>r</sup></li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
    <article id="September-29" class="note-item">
 +
        <h2>September 29</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="September-30" class="note-item">
 +
        <h2>September 30</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
 +
    <!--October-->
 +
 +
    <article id="October-1" class="note-item">
 +
        <h2>October-1</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-2" class="note-item">
 +
        <h2>October 2</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-3" class="note-item">
 +
        <h2>October 3</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-4" class="note-item">
 +
        <h2>October 4</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-5" class="note-item">
 +
        <h2>October 5</h2><hr>
 +
      <p class="note-title">Cultivation</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">For larva test</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-GS linker-lectin  A<sup>r</sup> </li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="October-6" class="note-item">
 +
        <h2>October 6</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-7" class="note-item">
 +
        <h2>October 7</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
 +
    <article id="October-8" class="note-item">
 +
        <h2>October 8</h2><hr>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">HLG-1</li>
 +
                      <li class="list">HLG-2</li>
 +
                      <li class="list">Hv1a-his </li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his </li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his </li>
 +
                      <li class="list">OAIP-lectin-his </li>
 +
                      <li class="list">BL</li>
 +
                      <li class="list">RG</li>
 +
                </ul>
 +
        </ul>
 +
 +
        <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">HLG-1</li>
 +
                      <li class="list">Hv1a-his </li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his </li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his </li>
 +
                      <li class="list">OAIP-lectin-his </li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="October-9" class="note-item">
 +
        <h2>October 9</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-10" class="note-item">
 +
        <h2>October 10</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-11" class="note-item">
 +
        <h2>October 11</h2><hr>
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">HLG-1</li>
 +
                      <li class="list">Hv1a-his </li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his </li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his </li>
 +
                      <li class="list">OAIP-lectin-his </li>
 +
                      <li class="list">RG</li>
 +
                </ul>
 +
        </ul>
 +
 +
        <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a(sonication)</li>
 +
                      <li class="list">Hv1a(purification)</li>
 +
                      <li class="list">Sf1a(sonication)</li>
 +
                      <li class="list">Sf1a(purification)</li>
 +
                      <li class="list">OAIP(sonication)</li>
 +
                      <li class="list">OAIP(purification)</li>
 +
                </ul>
 +
        </ul>
 +
 +
      <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">HLG</li>
 +
                      <li class="list">Hv1a-his </li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his </li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his </li>
 +
                      <li class="list">OAIP-lectin-his </li>
 +
                </ul>
 +
        </ul>
 +
 +
          <p class="note-title">Protein quantification Braford method</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">HLG(purification)</li>
 +
                      <li class="list">Hv1a-his(purification) </li>
 +
                      <li class="list">Hv1a-lectin-his(purification)</li>
 +
                      <li class="list">Sf1a-his(purification)</li>
 +
                      <li class="list">Sf1a-lectin-his(purification)</li>
 +
                      <li class="list">OAIP-his(purification) </li>
 +
                      <li class="list">OAIP-lectin-his(purification) </li>
 +
                      <li class="list">Hv1a-his(sonication)</li>
 +
                      <li class="list">OAIP-his(sonication) </li>
 +
                      <li class="list">OAIP-lectin-his(sonication) </li>
 +
                      <li class="list">HGL-his(sonication) </li>
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                      </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="October-12" class="note-item">
 +
        <h2>October 12</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-13" class="note-item">
 +
        <h2>October 13</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-14" class="note-item">
 +
        <h2>October 14</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-15" class="note-item">
 +
        <h2>October 15</h2><hr>
 +
        <p class="note-title">SDS-PAGE</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Check the purification protein.</li>
 +
              <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Hv1a-his</li>
 +
                      <li class="list">Hv1a-lectin-his</li>
 +
                      <li class="list">Sf1a-his</li>
 +
                      <li class="list">Sf1a-lectin-his</li>
 +
                      <li class="list">OAIP-his</li>
 +
                      <li class="list">OAIP-lectin-his</li>
 +
                </ul>
 +
        </ul>
 +
    </article>
 +
 +
    <article id="October-16" class="note-item">
 +
        <h2>October 16</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-17" class="note-item">
 +
        <h2>October 17</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-18" class="note-item">
 +
        <h2>October 18</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    <article id="October-19" class="note-item">
 +
        <h2>October 19</h2><hr>
 +
        <p class="note-title">None</p>
 +
    </article>
 +
    </section>
 
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Revision as of 17:26, 18 September 2018

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Wet Lab

July 1

None

July 2

None

July 3

none

July 4

None

July 5

Restore IDT in gblock fragment

  • Save DNA
  • Sample
    • Hv1a-his-Sf1a NO.98351612
    • Sf1a-his- OAIP NO.98351613
    • OAIP-his- Hv1a NO.98351614
    • Hv1a-lectin-his NO.98351615
    • Sf1a-lectin-his NO.98351616
    • OAIP-lectin-his NO.98351617

July 6

None

July 7

None

July 8

None

July 9

PCR of IDT gblock fragment

  • The IDT PCR amplify the insert gene and use phusion DNA polymerase (PFU) to improve the PCR accuracy rate.
  • Sample
    • Hv1a-his-Sf1a
    • Sf1a-his- OAIP
    • OAIP-his- Hv1a
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

Electrophoresis

  • Check IDT PCR product
  • Sample(7/9 PCR product)
    • Hv1a-his
    • Sf1a-his
    • OAIP-his
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

July 10

PCR of IDT gblock fragment

  • Amplify the DNA to revise the mistake.
  • Sample
    • Hv1a-his-Sf1a
    • Sf1a-his- OAIP
    • OAIP-his- Hv1a
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

Electrophoresis

  • Check IDT PCR product
  • Sample(7/10 PCR product)
    • Hv1a-his
    • Sf1a-his
    • OAIP-his
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

Gel extraction

  • For the purification of DNA, we cut the correct electrophoresis band to do gel extraction.
  • Sample
    • Hv1a-his
    • Sf1a-his
    • OAIP-his
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

Digestion

  • Digest the gene fragment(7/10) and pSB1C3 with EcoRI and PstI to ligase the gene fragment into the pSB1C3.
  • Sample
    • Hv1a-lectin-his 4 7.10 (EP)
    • Sf1a-his 1 7.10 (EP)
    • OAIP-his 2 7.10 (EP)
    • OAIP-lectin-his 5 7.10 (EP)
    • Hv1a-his 3 7.10 (EP)
    • Sf1a-lectin-his 6 7.10 (EP)
    • pSB1C3 (EP)

Electrophoresis

  • Electrophoresis of digested product to check the pSB1C3 digestion.
  • Sample
    • pSB1C3 (EP)

Ligation

  • Ligase DNA fragment into the pSB1C3
  • Sample digestion product
  • Sample
    • Hv1a-lectin-his 4 7.10 (EP)
    • Sf1a-his 1 7.10 (EP)
    • OAIP-his 2 7.10 (EP)
    • OAIP-lectin-his 5 7.10 (EP)
    • Hv1a-his 3 7.10 (EP)
    • Sf1a-lectin-his 6 7.10 (EP)
    • pSB1C3 (EP)

July 11

Transformation:

  • Transformation of ligated product to DH5α:
  • Sample
    • Sf1a-his 1 7.10 Cr
    • Hv1a-lectin-his 4 7.10 Cr
    • OAIP-his 2 7.10 Cr
    • OAIP-lectin-his 5 7.10 Cr
    • Hv1a-his 3 7.10 Cr
    • Sf1a-lectin-his 6 7.10 Cr

July 12

Electrophoresis

  • Electrophoresis of gel extraction product to check the concentration of gene to adjust the ligation protocol.
  • Sample
    • Hv1a-his
    • Sf1a-his
    • OAIP-his
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

Transformation:

  • Transformation of ligated product 20µL to BL21 competent cell 50µL
  • Sample
    • Sf1a-his 1 7.10 Cr
    • Hv1a-lectin-his 4 7.10 Cr
    • OAIP-his 2 7.10 Cr
    • OAIP-lectin-his 5 7.10 Cr
    • Hv1a-his 3 7.10 Cr
    • Sf1a-lectin-his 6 7.10 Cr

July 13

PCR

  • PCR of ligated products to amplify the insert gene.
  • Sample
    • Sf1a-his Cr 1~8
    • Hv1a-lectin-his Cr 1~8
    • OAIP-his Cr 1~8
    • OAIP-lectin-his Cr 1~8
    • Hv1a-his Cr 1~8
    • Sf1a-lectin-his Cr 1~8

July 14

PCR

  • PCR of ligated products to amplify the insert gene.
  • Sample
    • Sf1a-his Cr 1~4
    • Hv1a-lectin-his Cr 1~4
    • OAIP-his Cr 1~4
    • OAIP-lectin-his Cr 1~4
    • Hv1a-his Cr 1~4
    • Sf1a-lectin-his Cr 1~4

Electrophoresis

  • Electrophoresis of PCR products to check the insert gene base pair
  • Sample
    • Hv1a-his Cr 1~4
    • Sf1a-his Cr 1~4
    • OAIP-his Cr 1~4
    • Hv1a-lectin-his Cr 1~4
    • Sf1a-lectin-his Cr 1~4
    • OAIP-lectin-his Cr 1~4

Cultivation

  • Cultivation of ligated product.
  • Sample
    • Sf1a-his Cr 3
    • Hv1a-lectin-his Cr 4
    • OAIP-his Cr 1
    • OAIP-lectin-his Cr 3
    • Hv1a-his Cr 3
    • Sf1a-lectin-his Cr 4

July 15

PCR

  • PCR of ligated products to amplify the insert gene.
  • Sample
    • Sf1a-his Cr 1~8
    • Hv1a-lectin-his Cr 1~8
    • Hv1a-his Cr 1~6
    • Sf1a-lectin-his Cr 1~3

Electrophoresis

  • Electrophoresis of PCR products to check the insert gene base pair
  • Sample
    • Sf1a-his Cr 1~8
    • Hv1a-lectin-his Cr 1~8
    • Hv1a-his Cr 1~6
    • Sf1a-lectin-his Cr 1~3

Transformation

  • Transformation of Hv1a-lectin-his Cr, Hv1a-his Cr into BL21 again.

Miniprep

  • Purify plasmid which have insert gene.(7/14)
  • Sample
    • Hv1a-his Cr
    • Sf1a-his Cr
    • OAIP-his Cr
    • Hv1a-lectin-his Cr
    • Sf1a-lectin-his Cr
    • OAIP-lectin-his Cr

July 16

Digestion

  • Digest the plasmid (7/15) with EcoRI and PstI to check whether the mini have insert gene fragment.
  • Sample
    • Sf1a-his 7.15 (EP)
    • OAIP-his 7.15 (EP)
    • Hv1a-his 7.15 (EP)
    • Sf1a-lectin-his 7.15 (EP)
    • OAIP-lectin-his 7.15 (EP)
    • Hv1a-lectin-his 7.15 (EP)

Electrophoresis

  • Electrophoresis of digested product to check the band whether proper to the really base pair.
  • Sample(7/16 Digestion product)
    • Sf1a-his 7.15 (EP)
    • OAIP-his 7.15 (EP)
    • Hv1a-his 7.15 (EP)
    • Sf1a-lectin-his 7.15 (EP)
    • OAIP-lectin-his 7.15 (EP)
    • Hv1a-lectin-his 7.15 (EP)

PCR

  • PCR of ligated products to amplify the insert gene.
  • Sample
    • Hv1a-his Cr 1~11
    • Hv1a-lectin-his Cr 1~7

Electrophoresis

  • Electrophoresis of PCR products to check the insert gene base pair.
  • Sample
    • Hv1a-his Cr 1~11
    • Hv1a-lectin-his Cr 1~7

Cultivation

  • Cultivation of ligated product.
  • Sample
    • Sf1a-his Cr 1
    • Hv1a-his Cr 6
    • Hv1a-lectin-his Cr 1
    • Hv1a-lectin-his Cr 8
    • Sf1a-lectin-his Cr 1

July 17

Digestion

  • Digest the plasmid (7/15) with EcoRI and PstI to check whether the mini have insert gene fragment.
  • Sample
    • Sf1a-his 7.15 (EP)
    • Hv1a-his 7.15 (EP)
    • Sf1a-lectin-his 7.15 (EP)
    • Hv1a-lectin-his 7.15 (EP)

Electrophoresis

  • Electrophoresis of digested product to check the band whether proper to the really base pair.
  • Sample
    • Sf1a-his 7.15 (EP)
    • Hv1a-his 7.15 (EP)
    • Sf1a-lectin-his 7.15 (EP)
    • Hv1a-lectin-his 7.15 (EP)

Miniprep

  • Purify plasmid which have insert gene.(7/16)
  • Sample
    • Sf1a-his Cr 1
    • Sf1a-lectin-his Cr 1
    • Hv1a-his Cr 6
    • Hv1a-lectin-his Cr 1
    • Hv1a-lectin-his Cr 8

Cultivation

  • Cultivation of ligated product.(7/16)
  • Sample
    • Hv1a-his Cr 5
    • Hv1a-his Cr 7
    • Hv1a-his Cr 10
    • Hv1a-his Cr 11
    • Hv1a-lectin-his Cr 2
    • Hv1a-lectin-his Cr 3
    • Hv1a-lectin-his Cr 7

July 18

PRC of IDT gblock fragment:

  • Amplify the DNA to revise the mistake.
  • Sample
    • Hv1a-his-Sf1a NO.98351612
    • Sf1a-his-OAIP NO.98351613
    • OAIP-his-Hv1a NO.98351614
    • Hv1a-lectin-his NO.98351615
    • Sf1a-lectin-his NO.98351616
    • OAIP-lectin-his NO.98351617

Miniprep

  • Purify plasmid which have insert gene.(7/17)
  • Sample
    • Hv1a-his Cr 5
    • Hv1a-his Cr 7
    • Hv1a-his Cr 10
    • Hv1a-his Cr 11
    • Hv1a-lectin-his Cr 3
    • Hv1a-lectin-his Cr 7

Cultivation

  • Cultivation of ligated product.(7/17)
  • Sample
    • OAIP-lectin-his Cr 1
    • OAIP-lectin-his Cr 2

Digestion

  • Digest the plasmid (7/18) with EcoRI and PstI to check whether the mini have insert gene fragment.
  • Sample
    • Hv1a-his(EP)
    • Hv1a-his(EP)
    • Hv1a-his(EP)
    • Hv1a-his(EP)
    • Hv1a-lectin-his(EP)
    • Hv1a-lectin-his(EP)

Gel Electrophoresis of PCR-Amplified gBlocks

  • Sample
    • Hv1a-his
    • Sf1a-his
    • OAIP-his
    • Hv1a-lectin-his
    • sf1a-lectin-his
    • OAIP-lectin-his

Digestion

  • Digest the gene fragment(7/18) and pSB1A3 with EcoRI and PstI to ligase the gene fragment into the pSB1A3.
  • Sample
    • Sf1a-his 1 7.18 (EP)
    • Hv1a-lectin-his 4 7.18 (EP)
    • OAIP-his 2 7.18 (EP)
    • OAIP-lectin-his 5 7.18 (EP)
    • Hv1a-his 3 7.18 (EP)
    • Sf1a-lectin-his 6 7.18 (EP)
    • pSB1C3 (EP)

Electrophoresis

  • Electrophoresis of digested product to check the pSB1C3 digestion.
  • Sample7/18 Digestion product

Ligation

  • Ligase DNA fragment(7/18) into the pSB1C3
  • Sample
    • Sf1a-his 1 7.18 (EP)
    • Hv1a-lectin-his 4 7.18 (EP)
    • OAIP-his 2 7.18 (EP)
    • OAIP-lectin-his 5 7.18 (EP)
    • Hv1a-his 3 7.18 (EP)
    • Sf1a-lectin-his 6 7.18 (EP)
    • pSB1C3 (EP)

Electrophoresis

  • check the ligation product.
  • Sample7/18 Ligation product

July 19

Electrophoresis

  • Electrophoresis of ligated product.
  • Sample
    • Hv1a-his
    • Sf1a-his
    • OAIP-his
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

Transformation

  • Transformation of ligated product to DH5α.
  • Sample
    • Hv1a-his 7/18 Ar
    • Sf1a-his 7/18 Ar
    • OAIP-his 7/8 Ar
    • Hv1a-lectin-his 7/8 Ar
    • sf1a-lectin-his 7/8 Ar
    • OAIP-lectin-his 7/18 Ar

Miniprep

  • Purify plasmid which have insert gene.(7/18)
  • Sample
    • OAIP-lectin-his Cr 7/19 1
    • OAIP-lectin-his Cr 7/19 2

Digestion

  • Digest the plasmid (7/19) with EcoRI and PstI to check whether the mini have insert gene fragment.
  • Sample
    • Hv1a-lectin-his Cr 1 (EP)
    • Hv1a-lectin-his Cr 2 (EP)

Electrophoresis

  • Electrophoresis of ligated product.
  • Sample
    • Hv1a-lectin-his Cr 7/19 1
    • Hv1a-lectin-his Cr 7/19 2

Digestion

  • Digest the plasmid (7/18) with EcoRI and PstI to check whether the mini have insert gene fragment.
  • Sample
    • Hv1a-his Cr 1
    • Hv1a-his Cr 2
    • Hv1a-his Cr 3
    • Hv1a-his Cr 4
    • Hv1a-lectin-his Cr 1
    • Hv1a-lectin-his Cr 2

Electrophoresis

  • Electrophoresis of digestion product.
  • Sample
    • Hv1a-his Cr 1
    • Hv1a-his Cr 2
    • Hv1a-his Cr 3
    • Hv1a-his Cr 4
    • Hv1a-lectin-his Cr 1
    • Hv1a-lectin-his Cr 2

July 20

PCR

  • PCR of ligated products(7/19) to amplify the insert gene.
  • Sample
    • Hv1a-his Ar 1~4
    • Sf1a-his Ar 1~4
    • OAIP-his Ar 1~4
    • Hv1a-lectin-his Ar 1~4
    • Sf1a-lectin-his Ar 1~4
    • OAIP-lectin-his Ar 1~4

Digestion

  • Digest the gene fragment(7/18) and pSB1A3 with EcoRI and PstI to ligase the gene fragment into the pSB1A3.
  • Sample
    • Hv1a-his 7/18 Cr(EP)
    • Hv1a-his 7/18 Cr(EP)
    • Hv1a-his 7/18 Cr(EP)
    • Hv1a-his 7/18 Cr(EP)
    • Hv1a-lectin-his 7/18 Cr(EP)
    • Hv1a-lectin-his 7/18 Cr(EP)
    • OAIP-lectin-his 7/18 Cr(EP)

Electrophoresis

  • Electrophoresis of digestion product.
  • Sample
    • Hv1a-his 1~4 7/18(EP)
    • Hv1a-lectin-his 1, 2 7/18 (EP)
    • OAIP-lectin-his 7/19(EP)
    • Hv1a-his 3 7/18
    • Hv1a-lectin-his 2 7/18

Electrophoresis

  • Check insert gene in E.coli.
  • Sample
    • Hv1a-his 1-1~4
    • Hv1a-lectin-his 2-1~4
    • Sf1a-his3-1~4
    • Sf1a-lectin-his 4-1~4
    • OAIP-his 5-1~4
    • OAIP-lectin-his 6-1~4

Sequencing

  • determine insert gene’s Sequence.
  • Sample
    • Hv1a-his 3(The result is correct)
    • Hv1a-lectin-his 2
    • OAIP-lectin-his 1

Ligation

  • Ligase DNA fragment(7/18) into the pSB1A3
  • Sample digestion product
    • Hv1a-his 7/20 (EP)
    • Hv1a-lectin-his 7/20 (EP)
    • OAIP-lectin-his 7/20 (EP)

July 21

Cultivation

  • Cultivation of ligated product.(7/20)
  • Sample
    • Hv1a-his Ar 2~4
    • Sf1a-his Ar 1~4
    • OAIP-his Ar 1.3.4
    • Hv1a-lectin-his Ar 1~4
    • Sf1a-lectin-his Ar 1~4
    • OAIP-lectin-his Ar 1.3.4

Miniprep

  • Purify plasmid which have insert gene.(7/21)
  • Sample
    • Hv1a-his Ar 2~4
    • Sf1a-his Ar 1~4
    • OAIP-his Ar 1.3.4
    • Hv1a-lectin-his Ar 1~4
    • Sf1a-lectin-his Ar 1~4
    • OAIP-lectin-his Ar 1.3.4

Transformation

  • Transformation of ligated product(7/20) to BL21-rosetta-gami
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • OAIP-lectin-his Ar

July 22

Digestion

  • Digest the plasmid (7/21) with EcoRI and PstI to check whether the mini have insert gene fragment.
  • Sample
    • Hv1a-his Ar 2~4
    • Sf1a-his Ar 1~4
    • OAIP-his Ar 1.3.4
    • Hv1a-lectin-his Ar 1~4
    • Sf1a-lectin-his Ar 1~4
    • OAIP-lectin-his Ar 1.3.4

Electrophoresis

  • Electrophoresis of ligated product.
  • Sample
    • Hv1a-his Ar 2~4
    • Sf1a-his Ar 1~4
    • OAIP-his Ar 1.3.4
    • Hv1a-lectin-his Ar 1~4
    • Sf1a-lectin-his Ar 1~4
    • OAIP-lectin-his Ar 1.3.4

Sequencing

  • determine insert gene’s Sequence.
  • Sample
    • Hv1a-his (The result is correct)
    • Hv1a -his (The result is correct)
    • OAIP -his (The result is correct)

Digestion

  • Digest the plasmid (7/21) with EcoRI and PstI to check whether the mini have insert gene fragment.
  • Sample
    • Sf1a-his Ar 1~4(EP)
    • OAIP-his Ar 1.3.4(EP)
    • OAIP-lectin-his Ar 1.3.4(EP)

Digestion

  • Digest IDT gBlock fragment for ES because lectin have P Cut position.
  • Sample
    • Hv1a-lectin-his
    • OAIP-lectin-his
    • Sf1a-lectin-his
    • Psb1A3

Electrophoresis

  • Electrophoresis of ligated product.
  • Sample digestion product
    • Sf1a-his Ar 1~4(EP)
    • OAIP-his Ar 1.3.4(EP)
    • OAIP-lectin-his Ar 1.3.4(EP)

PRC of IDT gblock fragment

  • Amplify the DNA to revise the mistake.
  • Sample
    • Hv1a-his
    • Sf1a -his
    • OAIP-his
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

Electrophoresis

  • Electrophoresis of digested product to check insert gene.
  • Sample
    • Hv1a-lectin-his (ES)
    • Sf1a-lectin-his (ES)
    • OAIP-lectin-his (ES)
    • [pSB1A3] (ES)

Ligation

  • Ligase DNA fragment(7/18) into the pSB1A3
  • Sample
    • Hv1a-lectin-his (ES)
    • Sf1a-lectin-his (ES)
    • OAIP-lectin-his (ES)
    • [pSB1A3] (ES)

Transformation

  • Transformation of backbone plasmid into BL21-rosetta-gami check whether the competent cell is working.
  • we choose pSB1A3 to transformation because BL21-rosetta-gami has Cr and Kr.

July 23

Electrophoresis

  • After the PRC of IDT gBlock fragment, we electrophoresis samples to check the insert gene fragment.
  • Sample
    • Hv1a-his
    • Sf1a-his
    • OAIP-his
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

Digestion

  • There is a fragment in the snow drop lectin identification with PstI. Therefore, if sequence have snow drop lectin, we digested the gene fragment(7/22) and pSB1A3 with EcoRI and Spe l to ligase the gene fragment into the pSB1A3. Others still digested with EcoRI and PstI.
  • Sample
    • Hv1a-his 3 7/18 (EP)
    • Sf1a-his 1 7/8 (EP)
    • OAIP-his 2 7/18 (EP)
    • Hv1a-lectin-his 4 7/8 (ES)
    • Sf1a-lectin-his 6 7/18 (ES)
    • OAIP-lectin-his 5 7/18(ES)
    • pSB1A3 (EP)
    • pSB1A3 (ES)

Ligation

  • Ligase DNA fragment(7/23) into the pSB1A3
  • Sample
    • Hv1a-his 3 7/18 (EP)
    • Sf1a-his 1 7/8 (EP)
    • OAIP-his 2 7/18 (EP)
    • Hv1a-lectin-his 4 7/8 (ES)
    • Sf1a-lectin-his 6 7/18 (ES)
    • OAIP-lectin-his 5 7/18(ES)
    • pSB1A3 (EP)
    • pSB1A3 (ES)

Transformation

  • Transformation of ligated product.
  • Sample
    • Hv1a-lectin-his Ar 7/22
    • Sf1a-lectin-his Ar 7/22
    • OAIP-lectin-his Ar 7/22

July 24

PCR

  • PCR of ligated products(7/23) to amplify the insert gene.
  • Sample
    • Sf1a-lectin-his Ar 7/23 1~4
    • OAIP-lectin-his Ar 7/23 5~8
    • Hv1a-lectin-his Ar 7/23 9~12

Electrophoresis

  • Electrophoresis of PCR products
  • Sample
    • Sf1a-lectin-his Ar 7/3 1~4
    • OAIP-lectin-his Ar 7/23 5~8
    • Hv1a-lectin-his Ar 7/23 9~12

Cultivation

  • Cultivation of ligated product.(7/24)
  • Sample
    • Sf1a-lectin-his Ar 7/24 2.3
    • OAIP-lectin-his Ar 7/24 5.7
    • Hv1a-lectin-his Ar 7/24 9.12

July 25

Miniprep

  • Purify plasmid which have insert gene.(7/24)
  • Sample
    • Sf1a-lectin-his Ar 7/24 2.3
    • OAIP-lectin-his Ar 7/24 5.7
    • Hv1a-lectin-his Ar 7/24 9.12

Digestion

  • Digest the plasmid (7/25) with EcoRI and Spel to check whether the mini have insert gene fragment.
  • Sample
    • Sf1a-lectin-his Ar 2.3(ES)
    • OAIP-lectin-his Ar 5.7(ES)
    • Hv1a-lectin-his Ar 9.12(ES)

Electrophoresis

  • Electrophoresis of ligated product.
  • Sample
    • Sf1a-lectin-his Ar 2.3(ES)
    • OAIP-lectin-his Ar 5.7(ES)
    • Hv1a-lectin-his Ar 9.12(ES)

Sequencing

  • determine insert gene’s Sequence.
  • Sample
    • Hv1a-lectin-his (The result is correct)
    • Hv1a-lectin-his (The result is correct)
    • OAIP - lectin-his (The result is correct)

July 26

Prepare of CaCl2

  • for competent cell

Rosetta-gami competent cell prepare

  • for expression of disulfide bond.

Transformation

  • Transformation of backbone plasmid into BL21-rosetta-gami check whether the competent cell is working.
  • we choose pSB1A3 to transformation because BL21-rosetta-gami has Cr and Kr.
  • Only write colonies. NO RFP EXPRESSION

July 27

Transformation

  • Transformation of backbone plasmid into BL21-rosetta-gami check whether the competent cell is working.
  • we choose pSB1A3 to transformation because BL21-rosetta-gami has Cr and Kr.
  • Only write colonies. NO RFP EXPRESSION

Transformation

  • Transformation of plasmid which have correct sequence to DH5α.
  • Sample
    • Sf1a -his
    • OAIP-his
    • Hv1a -his

July 28

Cultivation

  • Cultivation of correct colonies. (7/27)
    • Sf1a -his Ar
    • OAIP-his Ar
    • Hv1a -his Ar

Test competent cell

  • BL21-rosetta-gami aren’t express RFP but write colonies. Therefore, we test competent cell whether they could save on Ar agar plate.
  • After cultivation about 14 hours, there are many write colonies. We think them may be contamination during preparing competent cell.

July 29

Miniprep

  • Purify plasmid which have correct insert gene(7/28) to conserve and transform to BL21-Rosetta-gami.
    • Sf1a -his Ar
    • OAIP-his Ar
    • Hv1a -his Ar

Transformation

  • Transformation of plasmid which have correct sequence to DH5α.
  • Sample
    • Sf1a-lectin-his 7/24 Ar
    • OAIP-lectin-his 7/24 Ar
    • Hv1a-lectin-his 7/24 Ar

July 30

Cultivation

  • Cultivation of correct colonies. (7/29)
    • Sf1a-lectin-his 7/29 Ar
    • OAIP-lectin-his 7/29 Ar
    • Hv1a-lectin-his 7/29 Ar

Miniprep

  • Purify plasmid which have correct insert gene(7/30) to conserve and transform to BL21-Rosetta-gami.
  • Sample
    • Sf1a-lectin-his 7/29 Ar
    • OAIP-lectin-his 7/29 Ar
    • Hv1a-lectin-his 7/29 Ar

Preparation of Competent cell

  • Rosetta-gami competent cell:
  • Prepare the bacteria that can make disulfide bond

July 31

None

August 1

Transformation

  • Transformation of pSB1A3 to test the competent cell work.
  • sample
    • PSB1A3

Transformation

  • Transformation of plasmid which have correct sequence to BL21-Rosetta-gami.
  • Sample
    • Hv1a-his
    • Sf1a-his
    • OAIP-his
    • Hv1a-lectin-his
    • Sf1a-lectin-his
    • OAIP-lectin-his

August 2

Transformation

  • Transformation of plasmid which have correct sequence to BL21-Rosetta-gami.
  • Sample
    • Sf1a-his 7/28 Ar
    • OAIP-his 7/28 Ar
    • Hv1a-lectin-his 7/29 Ar
    • Sf1a-lectin-his 7/29 Ar

August 3

Cultivation

  • Cultivation of BL21-Rosetta-gam colonies for IPTG induction.
  • sample
    • Hv1a-his 8/1 Ar
    • OAIP-lectin-his 8/1 Ar

Cultivation

  • Cultivation of correct colonies again for mini extraction.
  • Sample
    • Sf1a-his 7/27 Ar
    • OAIP-his 7/27 Ar
    • Hv1a-lectin-his 7/24 Ar
    • Sf1a-lectin-his 7/24 Ar

August 4

Miniprep

  • Purify plasmid which have correct insert gene(8/3) to conserve and transform to BL21-Rosetta-gami.
  • sample
    • OAIP-his 8/3 Ar
    • Sf1a-lectin-his 8/3 Ar
    • Hv1a-lectin-his 8/3 Ar

Cultivation

  • Cultivation of correct colonies again for mini extraction.
  • Sample
    • Sf1a-his 7/27 Ar

Transformation

  • Transformation of plasmid which have correct sequence to BL21-Rosetta-gami.
  • Sample
    • OAIP-his 8/3 Ar
    • Sf1a-lectin-his 8/3 Ar
    • Hv1a-lectin-his 8/3 Ar

IPTG induction

  • To find out the best condition for IPTG induction.
  • Sample
    • Hv1a-his 8/3 Ar
    • OAIP-lectin-his 8/3 Ar
  • Condition
    0μM, 250μM, 500μM, 750μM, 1000μM
  • Add IPTG concentration and test OD600
    0μM, 250μM, 500μM, 750μM, 1000μM

August 5

SDS-PAGE

  • Check the protein production after IPTG condition.
  • sample
    • Marker
    • OAIP-lectin-his 250μM 2hr
    • OAIP-lectin-his 500μM 2hr
    • OAIP-lectin-his 750μM 2hr
    • OAIP-lectin-his 1000μM 2hr
    • OAIP-lectin-his 0μM 4hr
    • OAIP-lectin-his 250μM 4hr
    • OAIP-lectin-his 500μM 4hr
    • OAIP-lectin-his 750μM 4hr
    • OAIP-lectin-his 1000μM 4hr
    • Marker
    • OAIP-lectin-his 250μM 6hr
    • OAIP-lectin-his 500μM 6hr
    • OAIP-lectin-his 750μM 6hr
    • OAIP-lectin-his 1000μM 6hr
    • OAIP-lectin-his 0μM 8hr
    • OAIP-lectin-his 250μM 8hr
    • OAIP-lectin-his 500μM 8hr
    • OAIP-lectin-his 750μM 8hr
    • OAIP-lectin-his 1000μM 8hr

August 6

None

August 7

None

August 8

None

August 9

None

August 10

None

August 11

None

August 12

None

August 13

None

August 14

Transformation

  • Transformation of plasmid which have correct sequence to BL21-Rosetta-gami.
  • Sample
    • Sf1a-his 7/29 Ar
    • OAIP-his 8/4 Ar
    • Hv1a-his 7/29 Ar
    • Sf1a-lectin-his 8/4 Ar
    • OAIP-lectin-his 7/30 Ar
    • Hv1a-lectin-his 8/4 Ar

PCR

  • PCR of transformation colonies to check the base pair.
  • Sample
    • Hv1a-his Ar 1~6
    • Hv1a-lectin-his Ar 1~6
    • Sf1a-lectin-his Ar 1~6
    • OAIP-lectin-his Ar 1~6

August 15

PCR

  • PCR of ligated products to amplify the insert gene.
  • Sample
    • Hv1a-his 8/14 Ar 1~6
    • Hv1a-lectin-his 8/14 Ar 1~6
    • Sf1a- lectin-his 8/14 Ar 1~6
    • Sf1a-lectin-his 8/4 Ar 1~6
    • OAIP-lectin-his 7/30 Ar 1~6
    • OAIP- lectin-his 8/14 Ar 1~6

Electrophoresis

  • Electrophoresis of PCR products to check the insert gene base pair
  • Sample
    • Hv1a-his 8/15 Ar 1~6
    • Hv1a-lectin-his 8/15 Ar 1~6
    • Sf1a- lectin-his 8/15 Ar 1~6
    • OAIP- lectin-his 8/15 Ar 1~6

Transformation

  • Transformation of plasmid which have correct sequence to BL21-Rosetta-gami.
  • Sample
    • Sf1a-his 7/29 Ar
    • OAIP-his 8/4 Ar

PCR

  • PCR of ligated products to amplify the insert gene.
  • Sample
    • Sf1a- lectin-his 8/15 Ar 1~6
    • OAIP- lectin-his 8/15 Ar 1~6

Electrophoresis

  • Electrophoresis of PCR products to check the insert gene base pair.
  • Sample
    • Sf1a- lectin-his 8/15 Ar 1~6
    • OAIP- lectin-his 8/15 Ar 1~6

SDS-PAGE

  • Check the production of IPTG induction (8/4) again.
  • No target protein expression.

August 16

Cultivation

  • Cultivation of BL21-Rosetta-gami colonies for IPTG induction.
  • Sample
    • Hv1a-his 8/15 Ar
    • Hv1a-lectin-his 8/15 Ar
    • Sf1a- lectin-his 8/15 Ar
    • OAIP-lectin-his 8/15 Ar

IPTG induction

  • To check the protein expression for IPTG induction.
  • Sample
    • Hv1a-his 8/16 Ar
    • Hv1a-lectin-his 8/16 Ar
    • Sf1a- lectin-his 8/16 Ar
    • OAIP-lectin-his 8/16 Ar
  • Condition
    0μM, 500μM
  • Add IPTG 500μM and test OD600 after 4hr.

Sonication and SDS-PAGE

  • Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.
  • Sample(8/16)
    • Hv1a-his 0μM 2hr
    • Hv1a-his 500μM 2hr
    • Sf1a-lectin-his 0μM 4hr
    • Sf1a-lectin-his 500μM 4hr
    • Hv1a-lectin-his 0μM 2hr
    • Hv1a-lectin-his 500μM 2hr
    • OAIP-lectin-his 0μM 4hr
    • OAIP-lectin-his 500μM 4hr
  • Failed. Do it again
  • Sample
    • Hv1a-his 0μM 2hr
    • Hv1a-his 500μM 2hr
    • Sf1a-lectin-his 0μM 4hr
    • Sf1a-lectin-his 500μM 4hr
    • Hv1a-lectin-his 0μM 2hr
    • Hv1a-lectin-his 500μM 2hr
    • OAIP-lectin-his 0μM 4hr
    • OAIP-lectin-his 500μM 4hr

August 17

IPTG induction

  • To check the protein expression for IPTG induction.
  • Sample
    • Hv1a-his 8/17 Ar
    • Hv1a-lectin-his
    • Sf1a- lectin-his
    • OAIP- lectin-his
  • Condition
    0μM, 500μM
  • Add IPTG 500μM and test OD600 after 4hr.

Sonication and SDS-PAGE

  • Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.
  • Sample
    • Hv1a-his 0μM 2hr
    • Hv1a-his 500μM 2hr
    • Sf1a-lectin-his 0μM 4hr
    • Sf1a-lectin-his 500μM 4hr
    • Hv1a-lectin-his 0μM 2hr
    • Hv1a-lectin-his 500μM 2hr
    • OAIP-lectin-his 0μM 4hr
    • OAIP-lectin-his 500μM 4hr

August 18

SDS-PAGE

  • To Check protein disulfide bond folding.
  • β-me make disulfide bond not folding.
  • Sample
    • Hv1a-lectin-his 0μM + β-me
    • Hv1a-lectin-his 500μM + β-me
    • Hv1a-lectin-his 500μM
    • Sf1a-lectin-his 0μM + β-me
    • Sf1a-lectin-his 500μM + β-me
    • Sf1a-lectin-his 500μM
    • OAIP-lectin-his 0μM + β-me
    • OAIP-lectin-his 500μM + β-me
    • OAIP-lectin-his 500μM

August 19

SDS-PAGE

  • To Check protein disulfide bond folding with the induction of control, Rosetta-gami stain.
  • β-me make disulfide bond not folding.
  • Sample(8/17)
    • Hv1a-lectin-his 0μM + β-me
    • Hv1a-lectin-his 500μM + β-me
    • Hv1a-lectin-his 500μM
    • Sf1a-lectin-his 0μM + β-me
    • Sf1a-lectin-his 500μM + β-me
    • Sf1a-lectin-his 500μM
    • Rosetta-gami + β-me
    • OAIP-lectin-his 0μM + β-me
    • OAIP-lectin-his 500μM + β-me
    • OAIP-lectin-his 500μM

August 20

Sonication and SDS-PAGE

  • Sonication and run SDS-PAGE to check peptides expression level after the IPTG induction.
  • Sample
    • Hv1a-his Ar
    • Hv1a-his Ar
    • Sf1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • Hv1a-lectin-his Ar
    • Hv1a-lectin-his Ar
    • OAIP-lectin-his Ar
    • OAIP-lectin-his Ar

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar 8/20
    • Hv1a-lectin-his Ar 8/20
    • Sf1a-lectin-hiss Ar 8/20
    • OAIP-lectin-his Ar 8/20

SDS-PAGE

  • To Check protein disulfide bond folding with the induction of control, Rosetta-gami stain.
  • Sample
    • Hv1a-his 8/19
    • Sf1a-his 8/19
    • OAIP-his 8/19

August 21

IPTG Induction and Sonication

  • To check the protein expression for IPTG induction. (Second round)
  • Sample
    • Hv1a-his Ar 8/20
    • Hv1a-lectin-his Ar 8/20
    • Sf1a-his Ar 8/17
    • Sf1a-lectin-his Ar 8/20
    • OAIP-his Ar 8/17
    • OAIP-lectin-his Ar 8/20
  • Condition
    0μM, 500μM
  • Add IPTG 500μM and test OD600 after 4hr.

August 22

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-lectin-his 8/20 Ar

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • BL21-Rosetta-gami competent cell
    • Cultivation 11 hours

IPTG Induction

  • Prepare for the purify the protein from sample BL21-Rosetta-gami colonies.
  • Sample
    • Sf1a-his Ar 8/20 Ar
  • Condition
    0μM, 1000μM
  • Add IPTG 500μM and test OD600 after 4hr.

Sonication

  • Sample
    • Hv1a-lectin-his 8/22
    • BL21-Rosetta-gami 8/22
    • Sf1a-lectin-his 0μM 8/22
    • Sf1a-lectin-his 1000μM 8/22

August 23

Protein purification(8/22-8/23)

  • Sample
    • Hv1a-lectin-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

Cultivation and IPTG induction

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar 8/20
    • Sf1a-lectin-his Ar 8/20
  • Add 1000μM IPTG and test OD600

Sonication

  • Sample
    • Hv1a-his Ar 8/23
    • Sf1a-lectin-his Ar 8/23

August 24

Protein purification(8/23-8/24)

  • Sample
    • Hv1a-lectin-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

Cultivation and IPTG induction

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Sf1a-his Ar 8/20
    • Sf1a-lectin-his Ar 8/20
  • Refresh to approximately OD600 0.7
  • Add 1000μM IPTG and test OD600
    • Sf1a-his: 0.966
    • Sf1a-lectin-his: 1.363
  • Certification, and add 2mL binding buffer and 200μM PMSF for sonication. Repeat three times.

Sonication

  • Sample
    • Sf1a-his Ar 8/24
    • Sf1a-lectin-his Ar 8/24

SDS-PAGE

  • Check the purification protein.
  • Sample
    • Hv1a-lectin-his(8/22-8/23)

August 25

Protein purification(8/24-8/25)

  • Sample
    • Hv1a-lectin-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample
    • Hv1a-lectin-his(8/23-8/24)

August 26

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar 8/15
    • Hv1a-lectin-his Ar 8/15
    • Sf1a-lectin-his Ar 8/15
    • OAIP-lectin-his Ar 8/15

Sonication

  • Sample
    • Sf1a-his Ar 8/24
    • Sf1a-lectin-his Ar 8/24

Protein purification

  • Sample
    • Hv1a-lectin-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Sf1a-his Ar 8/17
    • OAIP-his Ar 8/17

IPTG induction

  • Induce the mass protein expression for SDS-PAGE.
  • The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.
  • Sample
    • Hv1a-his Ar 8/26
    • Hv1a-lectin-his Ar 8/26
    • Sf1a-lectin-his Ar 8/26
    • OAIP-lectin-his Ar 8/26
  • Add 500μM IPTG
  • Take the following samples to go on the experience first. The others storage in 4℃ refrigerator.
    • Hv1a-his Ar 8/26
    • Hv1a-lectin-his Ar 8/26

SDS-PAGE

  • Check the protein in pellet or in solution.
  • Sample
    • Hv1a-his Ar 8/26(with urea)
    • Hv1a-his Ar 8/26
    • Hv1a-lectin-his Ar 8/26(with urea)
    • Hv1a-lectin-his Ar 8/26
  • Sonication and the supernatant is loading sample1
    • 10 seconds on, 40 seconds off and repeat six times.
  • Add binding buffer (with/without urea) again and the solution is loading sample 2
  • Sonication again and the supernatant is loading sample 3
    • 10 seconds on, 40 seconds off and repeat six times.
  • Add binding buffer (with/without urea) again and the solution is loading sample4
  • Loading sample
    • Hv1a-his (with urea) 1~4
    • Hv1a-his 1~4
    • Hv1a-lectin-his (with urea) 1~4
    • Hv1a-lectin-his 1~4

IPTG induction

  • Induce the mass protein expression for SDS-PAGE.
  • The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.
  • Sample
    • Hv1a-his Ar 8/26
    • Hv1a-lectin-his Ar 8/26
    • Sf1a-lectin-his Ar 8/26
    • OAIP-lectin-his Ar 8/26
  • Add 500μM IPTG and test OD600 to approximately 1.050

SDS-PAGE

  • Check the protein in pellet or in solution.
  • Sample
    • Hv1a-his Ar 8/26(with urea)
    • Hv1a-his Ar 8/26
    • Hv1a-lectin-his Ar 8/26(with urea)
    • Hv1a-lectin-his Ar 8/26
    • Sf1a-lectin-his Ar 8/26(with urea)
    • Sf1a-lectin-his Ar 8/26
    • OAIPlectin-his Ar 8/26(with urea)
    • OAIP-lectin-his Ar 8/26
  • Certificate the sample, add binding buffer (with/without urea), PMSF
  • Sonication and the supernatant is loading sample1
    • 10 seconds on, 40 seconds off and repeat six times.
  • Add binding buffer (with/without urea) again and the solution is loading sample 2
  • Sonication again and the supernatant is loading sample 3
    • 10 seconds on, 40 seconds off and repeat six times.
  • Add binding buffer (with/without urea) again and the solution is loading sample4
  • Loading sample
    • Hv1a-his (with urea) 1~4
    • Hv1a-his 1~4
    • Hv1a-lectin-his (with urea) 1~4
    • Hv1a-lectin-his 1~4
    • Sf1a-lectin-his (with urea) 1~4
    • Sf1a-lectin-his 1~4
    • OAIPlectin-his (with urea) 1~4
    • OAIP-lectin-his 1~4

August 27

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar

August 28

Protein purification

  • The buffer contains the urea
  • Sample(8/27)
    • Hv1a-his
    • Hv1a-lectin-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample(8/28)
    • Hv1a-his
    • Hv1a-lectin-his

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Sf1a-his Ar 8/17

August 29

Protein purification

  • The buffer contains the urea
  • Sample(8/28)
    • Sf1a-lectin-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample(8/29)
    • Sf1a-lectin-his

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Sf1a-his Ar 8/17

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar 8/15

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-lectin-his Ar 8/15
    • Sf1a-lectin-his Ar 8/15
    • OAIP-lectin-his Ar 8/15

August 30

IPTG induction

  • Induce the mass protein expression for SDS-PAGE.
  • The protein would form inclusion body. Therefore, it was devided into two parts, the urea and control.
  • Sample
    • Hv1a-lectin-his Ar 8/29
    • Sf1a-lectin-his Ar 8/29
    • OAIP-lectin-his Ar 8/29
  • Refresh to approximately OD600 0.6
  • Add 500μM IPTG and cultivate for 4hr

Protein purification

  • The buffer contains the urea
  • Sample(8/30)
    • Sf1a-lectin-his
    • Hv1a-lectin-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample(8/29)
    • Sf1a-lectin-his
    • Hv1a-lectin-his

PCR

  • amplify the insert gene
  • Sample(8/30)
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

Electrophoresis

  • Electrophoresis of PCR product
  • Sample(8/30 PCR product)
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

Digestion

  • Digest min for transformation
  • Sample
    • Hv1a-his digestion EP
    • Hv1a-lectin-his digestion ES
    • Sf1a-his digestion EP
    • Sf1a-lectin-his digestion ES
    • OAIP-his digestion EP
    • OAIP-lectin-his digestion ES

Transformation

  • Transformation PSB1C3 to DH5α
  • Sample
    • Trans PSB1C3 8/30
    • Trans PSB1C3 8/30

Urea test

  • Check whether protein in inclusion body or not.
  • Sample
    • Hv1a-his
    • Hv1a-his + Urea
    • Sf1a-his
    • Sf1a-his + Urea
    • OAIP-his
    • OAIP-his + Urea

August 31

Ligation

  • Ligase insert gene into the pSB1C3
  • Sample
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

Transformation

  • Transformation of ligated product to DH5α
  • Sample(8/31 Ligation product)
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

Dialysis and refolding

  • Dialysis and refolding for insoluble protein
  • Sample(8/8)
    • Hv1a-lectin-his (8/30)
    • Sf1a-lectin-his(8/30)
    • OAIP-lectin-his(8/30)
    • Hv1a-his(8/28)

SDS-PAGE

  • Check the purification protein.
  • Sample(8/30)
    • Sf1a-lectin-his
    • Hv1a-lectin-his

September 1

Protein purification

  • The buffer contains the urea
  • Sample(8/30)
    • OAIP-lectin-his
    • Hv1a-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample(8/30)
    • OAIP-lectin-his
    • Hv1a-his

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • OAIP-his Ar
    • Sf1a-his Ar

IPTG Induction

  • Prepare for the purify the protein from sample BL21-Rosetta-gami colonies.
  • Sample
    • Hv1a-his Ar
    • OAIP-his Ar
    • Sf1a-his Ar
  • Add IPTG 500μM and test OD600 after 4hr.

Sonication

  • Sample
    • Hv1a-his Ar
    • OAIP-his Ar
    • Sf1a-his Ar

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

Sonication

  • Sample
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

September 2

Protein purification

  • The buffer contains the urea
  • Sample
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

Protein quantification Braford method

  • Sample
    • Hv1a-his
    • Hv1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

Dialysis and refolding

  • Dialysis and refolding for insoluble protein
  • Sample
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

September 3

Protein purification

  • Sample
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

Protein quantification Braford method

  • The buffer contains the urea
  • Sample
    • Hv1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

Concentrate protein

  • Increase the concentration of protein
  • Sample
    • Hv1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

September 4

Protein purification

  • The buffer contains the urea
  • Sample
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

September 5

IPTG Induction

  • Prepare for the purify the protein from sample BL21-Rosetta-gami colonies.
  • Sample
    • Hv1a-his
    • Hv1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar
  • Add IPTG 500μM and test OD600 after 4hr.

Sonication

  • Sample
    • Hv1a-his
    • Hv1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

Protein purification

  • The buffer contains the urea
  • Sample
    • Hv1a-his
    • Hv1a-lectin-his
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

UV test

  • Test degradation rate
  • Sample
    • Hv1a-his
    • Hv1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

Concentrate protein

  • Increase the concentration of protein
  • Sample
    • Hv1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

Protein quantification Braford method

  • Sample
    • Hv1a-lectin-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar

September 6

Protein purification

  • The buffer contains the urea
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

Protein quantification Braford method

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

September 7

Protein purification

  • The buffer contains the urea
  • Sample
    • Sf1a-lectin-his Ar
    • OAIP-lectin-his Ar
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample
    • OAIP-lectin-his Ar
    • Sf1a-lectin-his Ar

UV test

  • Test degradation rate
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

Cultivation

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

Sonication

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

Protein quantification Braford method

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

September 8

Protein purification

  • The buffer contains the urea
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar

Cultivation and IPTG induction

  • Cultivation of sample BL21-Rosetta-gami colonies for protein purification.
  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar
  • Refresh to approximately OD600 0.7
  • Add 1000μM IPTG and test OD600
    • Hv1a-his:0.772
    • Hv1a-lectin-his:0.631
    • Sf1a-his:0.567
    • Sf1a-lectin-his:0.78
    • OAIP-his:0.851
    • OAIP-lectin-his:0.819
  • Certification, and add 2mL binding buffer and 200μM PMSF for sonication. Repeat three times.

Sonication

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

Sonication

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

Protein purification

  • The buffer contains the urea
  • Sample
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

September 9

Protein purification

  • The buffer contains the urea
  • Sample
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar
  • Collect：
    pass through solution, wash through solution(W1~3), elution solution(E1~10), filtered NaCl solution, and filtered ddH2O solution.

SDS-PAGE

  • Check the purification protein.
  • Sample
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

September 10

None

September 11

Dialysis protein

  • Dialysis and refolding for insoluble protein
  • Sample
    • Hv1a-his E1-E3
    • Hv1a-lectin-his E1-E5
    • Sf1a-his E1-E5
    • Sf1a-lectin-his E1-E4
    • OAIP-his E1-E3
    • OAIP-lectin-his E1-E2

September 12

Protein quantification Braford method

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

September 13

None

September 14

None

September 15

None

September 16

None

September 17

Dialysis protein

  • Dialysis and refolding for insoluble protein
  • Sample
    • Hv1a-his E2-E3
    • Hv1a-lectin-his E1-E6
    • Sf1a-his E1-E4
    • Sf1a-lectin-his E2-E4
    • OAIP-his E2-E4
    • OAIP-lectin-his E2-E3

September 18

None

September 19

None

September 20

None

September 21

None

September 22

Protein quantification Braford method

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

September 23

Dialysis protein

  • Remove urea
  • Sample
    • Hv1a-his E1-E3
    • Hv1a-lectin-his E1-E6
    • Sf1a-his E1-E4
    • Sf1a-lectin-his E1-E6
    • OAIP-his E1-E3
    • OAIP-lectin-his E1-E6

SDS-PAGE

  • Check the purification protein again
  • Sample
    • Hv1a-his
    • Hv1a-lectin-his (no β-me)
    • Sf1a-his
    • Sf1a-lectin-his (no β-me)
    • OAIP-his
    • OAIP-lectin-his (no β-me)

Electrophoresis

  • Electrophoresis of composite parts again
  • Sample(8/30 PCR product)
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

September 24

Electrophoresis

  • Electrophoresis of composite parts again
  • Sample(8/30 PCR product)
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

September 25

Protein quantification Braford method

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • Sf1a-lectin-his Ar
    • OAIP-his Ar
    • OAIP-lectin-his Ar

September 26

None

September 27

None

September 28

Protein quantification Braford method

  • Sample
    • Hv1a-his Ar
    • Hv1a-lectin-his Ar
    • Sf1a-his Ar
    • OAIP-his Ar

September 29

None

September 30

None

October-1

None

October 2

None

October 3

None

October 4

None

October 5

Cultivation

  • For larva test
  • Sample
    • Hv1a-GS linker-lectin Ar

October 6

None

October 7

None

October 8

Protein quantification Braford method

  • Sample
    • HLG-1
    • HLG-2
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his
    • BL
    • RG

Protein quantification Braford method

  • Sample
    • HLG-1
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

October 9

None

October 10

None

October 11

Protein quantification Braford method

  • Sample
    • HLG-1
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his
    • RG

SDS-PAGE

  • Check the purification protein.
  • Sample
    • Hv1a(sonication)
    • Hv1a(purification)
    • Sf1a(sonication)
    • Sf1a(purification)
    • OAIP(sonication)
    • OAIP(purification)

Protein quantification Braford method

  • Sample
    • HLG
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

Protein quantification Braford method

  • Sample
    • HLG(purification)
    • Hv1a-his(purification)
    • Hv1a-lectin-his(purification)
    • Sf1a-his(purification)
    • Sf1a-lectin-his(purification)
    • OAIP-his(purification)
    • OAIP-lectin-his(purification)
    • Hv1a-his(sonication)
    • OAIP-his(sonication)
    • OAIP-lectin-his(sonication)
    • HGL-his(sonication)
    • Hv1a-his
    • OAIP-lectin-his

October 12

None

October 13

None

October 14

None

October 15

SDS-PAGE

  • Check the purification protein.
  • Sample
    • Hv1a-his
    • Hv1a-lectin-his
    • Sf1a-his
    • Sf1a-lectin-his
    • OAIP-his
    • OAIP-lectin-his

October 16

None

October 17

None

October 18

None

October 19

None

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