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<!--Page_Content--> | <!--Page_Content--> | ||
<div class="container content"> | <div class="container content"> | ||
− | + | <div class="headstyle"> | |
+ | <h1 class="head">Notebook</h1> | ||
+ | </div> | ||
+ | <div class="righttitle"> | ||
+ | <h6 class="subtitle">Of Blood, Sweat and Tears</h6> | ||
+ | </div> | ||
<div class="navbar-example"> | <div class="navbar-example"> | ||
<div class="row"> | <div class="row"> | ||
<div class="col-2 side"> | <div class="col-2 side"> | ||
<div id="sidelist" class="list-group"> | <div id="sidelist" class="list-group"> | ||
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#Pasr_and_Pgada">P<sub>asr</sub> and P<sub>gadA</sub></a> |
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#pH_sensor_test">pH Sensor Test</a> |
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#CA">CA</a> |
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#RbcL">RbcL</a> |
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#RbcXS">RbcXS</a> |
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#PRK">PRK</a> |
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#Whole_Construction">Whole Construction</a> |
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#Total_Solution">Total Solution</a> |
<a class="list-group-item list-group-item-action" href="#"><i class="fa fa-arrow-up fa-1x" | <a class="list-group-item list-group-item-action" href="#"><i class="fa fa-arrow-up fa-1x" | ||
aria-hidden="true"></i> | aria-hidden="true"></i> | ||
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<div class="col-10"> | <div class="col-10"> | ||
<div data-spy="scroll" data-target="#sidelist" data-offset="0" class="scrollspy-example"> | <div data-spy="scroll" data-target="#sidelist" data-offset="0" class="scrollspy-example"> | ||
− | <div id=" | + | <div id="Pasr_and_Pgada"> |
− | < | + | <h3>Construction of P<sub>asr</sub> and P<sub>gadA</sub></h3></br> |
− | + | ||
<p class="pcontent">4/26~4/27</br> </p> | <p class="pcontent">4/26~4/27</br> </p> | ||
<ul> | <ul> | ||
− | <li>PCR | + | <li>PCR P<sub>asr</sub> and P<sub>gadA</sub> from <i>E. coli</i> MG1655 chromosome. Confirmed and extracted |
products by DNA gel. </li> | products by DNA gel. </li> | ||
</ul></br> | </ul></br> | ||
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<p class="pcontent">4/28</br> </p> | <p class="pcontent">4/28</br> </p> | ||
<ul> | <ul> | ||
− | <li>Digested the pSB1C3- | + | <li>Digested the pSB1C3-GFP plasmid. Ligase the plasmid with P<sub>asr</sub> / P<sub>gadA</sub> and |
RiboJ by PCR. Confirmed by DNA gel electrophoresis. Transformed the gene in DH5 | RiboJ by PCR. Confirmed by DNA gel electrophoresis. Transformed the gene in DH5 | ||
alpha and spread the bacteria on LB plates.</li> | alpha and spread the bacteria on LB plates.</li> | ||
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<p class="pcontent">5/9</br> </p> | <p class="pcontent">5/9</br> </p> | ||
<ul> | <ul> | ||
− | <li>Store the correct colonies in glycerol in - | + | <li>Store the correct colonies in glycerol in -80°C.</li> |
</ul></br> | </ul></br> | ||
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− | <div id=" | + | <div id="pH_sensor_test"> |
− | + | ||
<h3>The pH sensor test</h3></br> | <h3>The pH sensor test</h3></br> | ||
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<p class="pcontent">5/11</br> </p> | <p class="pcontent">5/11</br> </p> | ||
<ul> | <ul> | ||
− | <li>Pre-cultured the DH5 alpha-pSB1C3 - | + | <li>Pre-cultured the DH5 alpha-pSB1C3 -P<sub>asr</sub>-sfGFP strain.</li> |
</ul></br> | </ul></br> | ||
<p class="pcontent">5/12</br> </p> | <p class="pcontent">5/12</br> </p> | ||
<ul> | <ul> | ||
− | <li>Cultured the DH5 alpha-pSB1C3 - | + | <li>Cultured the DH5 alpha-pSB1C3 -P<sub>asr</sub>-sfGFP strain in M9 medium in different pH |
value. | value. | ||
− | Measured the | + | Measured the O.D. 600 value and fluorescence every hour.</li> |
</ul></br> | </ul></br> | ||
<p class="pcontent">5/18</br> </p> | <p class="pcontent">5/18</br> </p> | ||
<ul> | <ul> | ||
− | <li>Pre-cultured DH5 alpha-pSB1C3 | + | <li>Pre-cultured DH5 alpha-pSB1C3 –P<sub>gadA</sub>-RiboJ-sfGFP strain.</li> |
<li>Prepared the M9 medium in different pH value.</li> | <li>Prepared the M9 medium in different pH value.</li> | ||
</ul></br> | </ul></br> | ||
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<p class="pcontent">5/19</br> </p> | <p class="pcontent">5/19</br> </p> | ||
<ul> | <ul> | ||
− | <li>Cultured the DH5 alpha-pSB1C3 | + | <li>Cultured the DH5 alpha-pSB1C3 –P<sub>gadA</sub>-RiboJ-sfGFP strain in M9 medium in different |
pH | pH | ||
− | value. Measured the | + | value. Measured the O.D. 600 value and fluorescence every hour.</li> |
</ul></br> | </ul></br> | ||
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<p class="pcontent">7/29</br> </p> | <p class="pcontent">7/29</br> </p> | ||
<ul> | <ul> | ||
− | <li>Pre-culture the BL21(DE3)-pSB1C3 | + | <li>Pre-culture the BL21(DE3)-pSB1C3 –P<sub>asr</sub>-sfGFP strain</li> |
</ul></br> | </ul></br> | ||
<p class="pcontent">7/30</br> </p> | <p class="pcontent">7/30</br> </p> | ||
<ul> | <ul> | ||
− | <li>Cultured the BL21(DE3)-pSB1C3 | + | <li>Cultured the BL21(DE3)-pSB1C3 –P<sub>asr</sub> -sfGFP strain on 96 well plate in different pH |
value. Measure the fluorescence every 5 minutes in 30 minutes.</li> | value. Measure the fluorescence every 5 minutes in 30 minutes.</li> | ||
</ul></br> | </ul></br> | ||
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<p class="pcontent">9/4</br> </p> | <p class="pcontent">9/4</br> </p> | ||
<ul> | <ul> | ||
− | <li>Pre-culture the BL21(DE3)-pSB1C3 | + | <li>Pre-culture the BL21(DE3)-pSB1C3 –P<sub>gadA</sub>-RiboJ-sfGFP strain</li> |
</ul></br> | </ul></br> | ||
<p class="pcontent">9/5</br> </p> | <p class="pcontent">9/5</br> </p> | ||
<ul> | <ul> | ||
− | <li>Cultured the BL21(DE3)-pSB1C3 | + | <li>Cultured the BL21(DE3)-pSB1C3 –P<sub>gadA</sub>-RiboJ-sfGFP strain strain on 96 well plate in |
different pH value. Measure the fluorescence every 5 minutes in 30 minutes.</li> | different pH value. Measure the fluorescence every 5 minutes in 30 minutes.</li> | ||
</ul></br> | </ul></br> | ||
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− | <div id=" | + | <div id="CA"> |
− | + | <h3>Construction of CA</h3></br> | |
<p class="pcontent">7/2</br> </p> | <p class="pcontent">7/2</br> </p> | ||
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<p class="pcontent">7/12</br> </p> | <p class="pcontent">7/12</br> </p> | ||
<ul> | <ul> | ||
− | <li>Transformed the pSB1C3- | + | <li>Transformed the pSB1C3-P<sub>T7</sub>-CA into DH5 alpha. </li> |
</ul></br> | </ul></br> | ||
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− | <div id=" | + | <div id="RbcL"> |
− | + | ||
<h3>Construction of RbcL</h3></br> | <h3>Construction of RbcL</h3></br> | ||
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<p class="pcontent">8/1</br> </p> | <p class="pcontent">8/1</br> </p> | ||
<ul> | <ul> | ||
− | <li>Ligate the RbcL with | + | <li>Ligate the RbcL with P<sub>LacI</sub>.</li> |
− | <li>Transformed the pSB1C3- | + | <li>Transformed the pSB1C3-P<sub>T7</sub>-RbcL and pSB1C3-P<sub>LacI</sub>-RbcL into DH5 alpha.</li> |
</ul></br> | </ul></br> | ||
<p class="pcontent">8/3 </br> </p> | <p class="pcontent">8/3 </br> </p> | ||
<ul> | <ul> | ||
− | <li>Confirmed the pSB1C3- | + | <li>Confirmed the pSB1C3-P<sub>T7</sub>-RbcL construct by colony PCR.</li> |
<li>Sent the gene to sequencing</li> | <li>Sent the gene to sequencing</li> | ||
</ul></br> | </ul></br> | ||
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− | <div id=" | + | <div id="RbcXS"> |
− | + | ||
<h3>Construction of RbcXS</h3></br> | <h3>Construction of RbcXS</h3></br> | ||
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− | <div id=" | + | <div id="PRK"> |
− | + | ||
<h3>Construction of PRK</h3></br> | <h3>Construction of PRK</h3></br> | ||
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<p class="pcontent">8/1</br> </p> | <p class="pcontent">8/1</br> </p> | ||
<ul> | <ul> | ||
− | <li>Ligated PRK with | + | <li>Ligated PRK with P<sub>LacI</sub> and P<sub>T7</sub>.</li> |
<li>Transformed into DH5 alpha. | <li>Transformed into DH5 alpha. | ||
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− | <div id=" | + | <div id="Whole_Construction"> |
− | + | ||
<h3>Whole construction</h3></br> | <h3>Whole construction</h3></br> | ||
<p class="pcontent">8/8</br> </p> | <p class="pcontent">8/8</br> </p> | ||
<ul> | <ul> | ||
− | <li>Ligated | + | <li>Ligated P<sub>T7</sub>-CA with P<sub>LacI</sub>-PRK on pSB1C3</li> |
</ul></br> | </ul></br> | ||
<p class="pcontent">8/9</br> </p> | <p class="pcontent">8/9</br> </p> | ||
<ul> | <ul> | ||
− | <li>Ligated | + | <li>Ligated P<sub>T7</sub>-RbcL with P<sub>T7</sub>-RbcXS on pSB1C3</li> |
</ul></br> | </ul></br> | ||
<p class="pcontent">8/28</br> </p> | <p class="pcontent">8/28</br> </p> | ||
<ul> | <ul> | ||
− | <li>Inserted | + | <li>Inserted P<sub>T7</sub>-CA with P<sub>LacI</sub>-PRK into pSB3K3</li> |
</ul></br> | </ul></br> | ||
</div> | </div> | ||
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− | <div id=" | + | <div id="Total_Solution"> |
− | + | ||
<h3>SDS PAGE, Functional test and Total solution</h3></br> | <h3>SDS PAGE, Functional test and Total solution</h3></br> | ||
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<p class="pcontent">9/2</br> </p> | <p class="pcontent">9/2</br> </p> | ||
<ul> | <ul> | ||
− | <li> Ran SDS-PAGE of pSB1C3- | + | <li> Ran SDS-PAGE of pSB1C3-P<sub>T7</sub>-RbcL-P<sub>T7</sub>-RbcXS and pSB3K3-P<sub>LacI</sub>-PRK-P<sub>T7</sub>-CA</li> |
</ul></br> | </ul></br> | ||
<p class="pcontent">9/3</br> </p> | <p class="pcontent">9/3</br> </p> | ||
<ul> | <ul> | ||
− | <li> Transformed pSB1C3- | + | <li> Transformed pSB1C3-P<sub>T7</sub>-RbcL-P<sub>T7</sub>-RbcXS and pSB3K3-P<sub>LacI</sub>-PRK-P<sub>T7</sub>-CA into W3110</li> |
</ul></br> | </ul></br> | ||
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$(document).ready(function () { | $(document).ready(function () { | ||
$(window).scroll(function () { | $(window).scroll(function () { | ||
− | if ($(this).scrollTop() >= | + | if ($(this).scrollTop() >= 500) { |
var position = $("#sidelist").position(); | var position = $("#sidelist").position(); | ||
if (position == undefined) {} else { | if (position == undefined) {} else { |
Latest revision as of 20:13, 17 October 2018