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

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.menu {
 
.menu {
margin-left:6vw;
+
margin-left:10%;
        margin-top:10vh;
+
  margin-top:10vh;
 
width: 35%;
 
width: 35%;
 
min-width: 400px;
 
min-width: 400px;
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.notebook{
 
.notebook{
     margin-left:48vw;
+
     margin-left:55%;
 
     margin-top:-450px;
 
     margin-top:-450px;
 
     margin-bottom:30px;
 
     margin-bottom:30px;
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     <article id="July-1" class="note-item note_active">
 
     <article id="July-1" class="note-item note_active">
 
         <h2>July 1</h2><hr>
 
         <h2>July 1</h2><hr>
         <p class="note-title">None</p>
+
         <p class="note-title">Cloning</p>
 +
        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
            <li class="list">Received and resuspended of the IDT basic part</li>
 +
            <li class="list">Sample</li>
 +
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                      <li class="list">Enterocin A</li>
 +
                      <li class="list">Enterocin B</li>
 +
                      <li class="list">Enterocin 96</li>
 +
                      <li class="list">Bovicin HJ50</li>
 +
                      <li class="list">Duracin TW49M</li>
 +
                      <li class="list">Lacticin Z</li>
 +
                      <li class="list">Leucocyclicin Q</li>
 +
                      <li class="list">Subtilosin</li>
 +
                </ul>
 +
        </ul>
 
     </article>
 
     </article>
 
     <article id="July-2" class="note-item">
 
     <article id="July-2" class="note-item">
Line 679: Line 693:
 
     <article id="July-3" class="note-item">
 
     <article id="July-3" class="note-item">
 
         <h2>July 3</h2><hr>
 
         <h2>July 3</h2><hr>
         <p class="note-title">none</p>
+
         <p class="note-title">Growth curve exp</p>
 +
        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
            <li class="list">Observe the growth population of Bacillis subtilis in different temperature Condition: 37°C, 32°C, 27°C</li>
 +
        </ul>
 
     </article>
 
     </article>
 
     <article id="July-4" class="note-item">
 
     <article id="July-4" class="note-item">
Line 688: Line 705:
 
     <article id="July-5" class="note-item">
 
     <article id="July-5" class="note-item">
 
         <h2>July 5</h2><hr>
 
         <h2>July 5</h2><hr>
         <p class="note-title">Restore IDT in gblock fragment</p>
+
         <p class="note-title">None</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>
  
Line 713: Line 718:
 
     <article id="July-8" class="note-item">
 
     <article id="July-8" class="note-item">
 
         <h2>July 8</h2><hr>
 
         <h2>July 8</h2><hr>
         <p class="note-title">None</p>
+
         <p class="note-title">Growth curve exp</p>
 +
        <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
            <li class="list">Observe the growth population of Bacillis subtilis in different pH Condition: pH = 4, 5, 6, 7, 8</li>
 +
        </ul>
 
     </article>
 
     </article>
  
 
     <article id="July-9" class="note-item">
 
     <article id="July-9" class="note-item">
 
         <h2>July 9</h2><hr>
 
         <h2>July 9</h2><hr>
         <p class="note-title">PCR of IDT gblock fragment</p>
+
         <p class="note-title">None</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>
  
 
     <article id="July-10" class="note-item">
 
     <article id="July-10" class="note-item">
 
         <h2>July 10</h2><hr>
 
         <h2>July 10</h2><hr>
         <p class="note-title">PCR of IDT gblock fragment</p>
+
         <p class="note-title">None</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>
  
 
     <article id="July-11" class="note-item">
 
     <article id="July-11" class="note-item">
 
         <h2>July 11</h2><hr>
 
         <h2>July 11</h2><hr>
         <p class="note-title">Transformation:</p>
+
         <p class="note-title">None</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>
  
 
     <article id="July-12" class="note-item">
 
     <article id="July-12" class="note-item">
 
         <h2>July 12</h2><hr>
 
         <h2>July 12</h2><hr>
         <p class="note-title">Electrophoresis</p>
+
         <p class="note-title">None</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>
  
 
     <article id="July-13" class="note-item">
 
     <article id="July-13" class="note-item">
 
         <h2>July 13</h2><hr>
 
         <h2>July 13</h2><hr>
         <p class="note-title">PCR</p>
+
         <p class="note-title">None</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>
  
 
     <article id="July-14" class="note-item">
 
     <article id="July-14" class="note-item">
 
         <h2>July 14</h2><hr>
 
         <h2>July 14</h2><hr>
         <p class="note-title">PCR</p>
+
         <p class="note-title">None</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>
  
 
     <article id="July-15" class="note-item">
 
     <article id="July-15" class="note-item">
 
         <h2>July 15</h2><hr>
 
         <h2>July 15</h2><hr>
         <p class="note-title">PCR</p>
+
         <p class="note-title">Expression</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">repared competent cell E. coli ER2566</li>
              <li class="list">Sample</li>
+
          </ul>
                <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>
+
       <p class="note-title">Growth curve exp</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Observe the growth population of Bacillis subtilis in different salinity Condition: 0.17M, 0.25M, 0.5M, 0.75M, 1.0M</li>
 
               <li class="list">Sample</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>
 
           </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>
  
 
     <article id="July-16" class="note-item">
 
     <article id="July-16" class="note-item">
 
         <h2>July 16</h2><hr>
 
         <h2>July 16</h2><hr>
         <p class="note-title">Digestion</p>
+
         <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Amplify the insert gene (Pfu PCR)</li>
 +
              <li class="list">Electrophoresis (To check PCR products)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Lacticin Z + pTXB1</li>
                       <li class="list">OAIP-his 7.15 (EP)</li>
+
                       <li class="list">Bovincin HJ50+ pTXB1</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>
        </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>
  
 
     <article id="July-17" class="note-item">
 
     <article id="July-17" class="note-item">
 
         <h2>July 17</h2><hr>
 
         <h2>July 17</h2><hr>
         <p class="note-title">Digestion</p>
+
         <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Digestion of PCR products (Including insert and backbone pTXB1)</li>
 +
              <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Lacticin Z + pTXB1</li>
                       <li class="list">Hv1a-his 7.15 (EP)</li>
+
                       <li class="list">Bovincin HJ50+ pTXB1</li>
                      <li class="list">Sf1a-lectin-his 7.15 (EP)</li>
+
                      <li class="list">Hv1a-lectin-his 7.15 (EP)</li>
+
 
                 </ul>
 
                 </ul>
        </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>
 
     <article id="July-18" class="note-item">
 
     <article id="July-18" class="note-item">
 
         <h2>July 18</h2><hr>
 
         <h2>July 18</h2><hr>
       <p class="note-title">PRC of IDT gblock fragment:</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Transformation of ligation products (Transform into E. coli DH5α)</li>
 +
              <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li>
 +
              <li class="list">Electrophoresis (To check PCR products)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Lacticin Z + pTXB1</li>
                       <li class="list">Sf1a-his-OAIP NO.98351613</li>
+
                       <li class="list">Bovincin HJ50+ pTXB1</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>
        </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>
  
 
     <article id="July-19" class="note-item">
 
     <article id="July-19" class="note-item">
 
         <h2>July 19</h2><hr>
 
         <h2>July 19</h2><hr>
       <p class="note-title">Electrophoresis</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
               <li class="list">Electrophoresis of ligated product.</li>
+
               <li class="list">Cultivation</li>
 +
              <li class="list">Miniprep (Purify Plasmid)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
                       <li class="list">Hv1a-his</li>
+
                       <li class="list">Lacticin Z + pTXB1</li>
                       <li class="list">Sf1a-his</li>
+
                       <li class="list">Bovincin HJ50+ pTXB1</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>
        </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>
  
 
     <article id="July-20" class="note-item">
 
     <article id="July-20" class="note-item">
 
         <h2>July 20</h2><hr>
 
         <h2>July 20</h2><hr>
       <p class="note-title">PCR</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">PurposeL: Sequencing plasmid</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Lacticin Z + pTXB1</li>
                       <li class="list">Sf1a-his A<sup>r</sup> 1~4</li>
+
                       <li class="list">Bovincin HJ50+ pTXB1</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>
 
         </ul>
 
         </ul>
Line 1,388: Line 840:
 
         <h2>July 21</h2><hr>
 
         <h2>July 21</h2><hr>
 
       <p class="note-title">Cultivation</p>
 
       <p class="note-title">Cultivation</p>
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+
    </article>
              <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">
 
     <article id="July-22" class="note-item">
 
         <h2>July 22</h2><hr>
 
         <h2>July 22</h2><hr>
       <p class="note-title">Digestion</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Amplify the insert gene (Pfu PCR)</li>
 +
              <li class="list">Electrophoresis (To check PCR products)</li>
 +
              <li class="list">Digestion of PCR products (Including insert and backbone pTXB1)</li>
 +
              <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Leucocyclicin Q + pTXB1</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>
        </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>
 
         </ul>
 
     </article>
 
     </article>
Line 1,553: Line 859:
 
     <article id="July-23" class="note-item">
 
     <article id="July-23" class="note-item">
 
         <h2>July 23</h2><hr>
 
         <h2>July 23</h2><hr>
       <p class="note-title">Electrophoresis</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Transformation of ligation products (Transform into E. coli DH5α)</li>
 +
              <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li>
 +
              <li class="list">Electrophoresis (To check PCR products)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
                       <li class="list">Hv1a-his</li>
+
                       <li class="list">Leucocyclicin Q + pTXB1</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>
 
         </ul>
 
         </ul>
Line 1,613: Line 873:
 
     <article id="July-24" class="note-item">
 
     <article id="July-24" class="note-item">
 
         <h2>July 24</h2><hr>
 
         <h2>July 24</h2><hr>
       <p class="note-title">PCR</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Cultivation</li>
 +
              <li class="list">Miniprep (Purify Plasmid)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Leucocyclicin Q + pTXB1</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>
 
         </ul>
 
         </ul>
Line 1,650: Line 886:
 
     <article id="July-25" class="note-item">
 
     <article id="July-25" class="note-item">
 
         <h2>July 25</h2><hr>
 
         <h2>July 25</h2><hr>
       <p class="note-title">Miniprep</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Sequencing plasmid</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Leucocyclicin Q + pTXB1</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>
 
         </ul>
 
         </ul>
Line 1,698: Line 898:
 
     <article id="July-26" class="note-item">
 
     <article id="July-26" class="note-item">
 
         <h2>July 26</h2><hr>
 
         <h2>July 26</h2><hr>
       <p class="note-title">Prepare of CaCl<sub>2</sub></p>
+
       <p class="note-title">None</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>
  
 
     <article id="July-27" class="note-item">
 
     <article id="July-27" class="note-item">
 
         <h2>July 27</h2><hr>
 
         <h2>July 27</h2><hr>
         <p class="note-title">Transformation</p>
+
         <p class="note-title">None</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>
  
 
     <article id="July-28" class="note-item">
 
     <article id="July-28" class="note-item">
 
         <h2>July 28</h2><hr>
 
         <h2>July 28</h2><hr>
         <p class="note-title">Cultivation</p>
+
         <p class="note-title">None</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>
  
 
     <article id="July-29" class="note-item">
 
     <article id="July-29" class="note-item">
 
         <h2>July 29</h2><hr>
 
         <h2>July 29</h2><hr>
       <p class="note-title">Miniprep</p>
+
       <p class="note-title">Expression</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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>
+
               <li class="list">Transformation (Transform correct plasmid into E. coli ER2566)</li>
 
                     <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                     <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">Leucocyclicin Q + pTXB1</li>
                        <li class="list">OAIP-his A<sup>r</sup></li>
+
                        <li class="list">Hv1a -his A<sup>r</sup></li>
+
 
                     </ul>
 
                     </ul>
 
           </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>
  
 
     <article id="July-30" class="note-item">
 
     <article id="July-30" class="note-item">
 
         <h2>July 30</h2><hr>
 
         <h2>July 30</h2><hr>
       <p class="note-title">Cultivation</p>
+
       <p class="note-title">None</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>
 
     <article id="July-31" class="note-item">
 
     <article id="July-31" class="note-item">
Line 1,821: Line 935:
 
     <article id="August-1" class="note-item">
 
     <article id="August-1" class="note-item">
 
         <h2>August 1</h2><hr>
 
         <h2>August 1</h2><hr>
       <p class="note-title">Transformation</p>
+
       <p class="note-title">None</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>
  
 
     <article id="August-2" class="note-item">
 
     <article id="August-2" class="note-item">
 
         <h2>August 2</h2><hr>
 
         <h2>August 2</h2><hr>
       <p class="note-title">Transformation</p>
+
       <p class="note-title">None</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>
  
 
     <article id="August-3" class="note-item">
 
     <article id="August-3" class="note-item">
 
         <h2>August 3</h2><hr>
 
         <h2>August 3</h2><hr>
       <p class="note-title">Cultivation</p>
+
       <p class="note-title">None</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>
  
 
     <article id="August-4" class="note-item">
 
     <article id="August-4" class="note-item">
 
         <h2>August 4</h2><hr>
 
         <h2>August 4</h2><hr>
       <p class="note-title">Miniprep</p>
+
       <p class="note-title">None</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>
  
 
     <article id="August-5" class="note-item">
 
     <article id="August-5" class="note-item">
 
         <h2>August 5</h2><hr>
 
         <h2>August 5</h2><hr>
       <p class="note-title">SDS-PAGE</p>
+
       <p class="note-title">Expression</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Purepose: Transformation (Transform correct plasmid into E. coli ER2566)</li>
 
               <li class="list">sample</li>
 
               <li class="list">sample</li>
 
                     <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                     <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
                         <li class="list">Marker</li>
+
                         <li class="list">Leucocyclicin Q + pTXB1</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>
 
           </ul>
 
           </ul>
Line 1,969: Line 968:
 
     <article id="August-6" class="note-item">
 
     <article id="August-6" class="note-item">
 
         <h2>August 6</h2><hr>
 
         <h2>August 6</h2><hr>
         <p class="note-title">None</p>
+
         <p class="note-title">Expression</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purepose: Cultivation (Cultivation of E. coli ER2566 colonies for IPTG induction)</li>
 +
              <li class="list">Purepose: IPTG Induction (To test the OD levels of E. coli ER2566 to induce)Condition: OD = 0.5, 1.3</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Leucocyclicin Q + pTXB1</li>
 +
                    </ul>
 +
          </ul>
 
     </article>
 
     </article>
 
     <article id="August-7" class="note-item">
 
     <article id="August-7" class="note-item">
 
         <h2>August 7</h2><hr>
 
         <h2>August 7</h2><hr>
         <p class="note-title">None</p>
+
         <p class="note-title">Expression</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purepose: SDS PAGE (To check the protein production after induction)</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Leucocyclicin Q + pTXB1</li>
 +
                    </ul>
 +
          </ul>
 
     </article>
 
     </article>
 
     <article id="August-8" class="note-item">
 
     <article id="August-8" class="note-item">
Line 1,989: Line 1,003:
 
     <article id="August-11" class="note-item">
 
     <article id="August-11" class="note-item">
 
         <h2>August 11</h2><hr>
 
         <h2>August 11</h2><hr>
         <p class="note-title">None</p>
+
         <p class="note-title">Cloning</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purepose: Amplify the insert gene (Pfu PCR)</li>
 +
              <li class="list">Purepose: Electrophoresis (To check PCR products)</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Enterocin A</li>
 +
                        <li class="list">Enterocin B</li>
 +
                        <li class="list">Enterocin 96 </li>
 +
                        <li class="list">Duracin TW49M </li>
 +
                        <li class="list">Subtilosin</li>
 +
                    </ul>
 +
          </ul>
 
     </article>
 
     </article>
 
     <article id="August-12" class="note-item">
 
     <article id="August-12" class="note-item">
 
         <h2>August 12</h2><hr>
 
         <h2>August 12</h2><hr>
         <p class="note-title">None</p>
+
         <p class="note-title">Cloning</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purepose: Digestion of PCR products (Including insert and backbone pTXB1)</li>
 +
              <li class="list">Purepose: Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li>
 +
              <li class="list">Purepose: Transformation of ligation products (Transform into E. coli DH5α)</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Enterocin A + pTXB1</li>
 +
                        <li class="list">Enterocin B + pTXB1</li>
 +
                        <li class="list">Enterocin 96  + pTXB1</li>
 +
                        <li class="list">Duracin TW49M  + pTXB1</li>
 +
                        <li class="list">Subtilosin + pTXB1</li>
 +
                    </ul>
 +
          </ul>
 +
        <p class="note-title">Growth curve</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purepose: Observe the growth of Bacillus subtilis in different temperature, pH, salinityCondition:</li>
 +
              <li class="list">1.Temp: 37°C , pH:7 , salinity: 0.17M</li>
 +
              <li class="list">2.Temp: 30°C , pH:9 , salinity: 0.5M</li>
 +
              <li class="list">3.Temp: 25°C , pH:5 , salinity: 0.25M</li>
 +
          </ul>
 
     </article>
 
     </article>
 
     <article id="August-13" class="note-item">
 
     <article id="August-13" class="note-item">
 
         <h2>August 13</h2><hr>
 
         <h2>August 13</h2><hr>
         <p class="note-title">None</p>
+
         <p class="note-title">Cloning</p>
 +
          <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
              <li class="list">Purepose: Taq PCR (PCR of ligation products to amplify insert gene)</li>
 +
              <li class="list">Purepose: Electrophoresis (To check PCR products)</li>
 +
              <li class="list">sample</li>
 +
                    <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 +
                        <li class="list">Enterocin A + pTXB1</li>
 +
                        <li class="list">Enterocin B + pTXB1</li>
 +
                        <li class="list">Enterocin 96  + pTXB1</li>
 +
                        <li class="list">Duracin TW49M  + pTXB1</li>
 +
                        <li class="list">Subtilosin + pTXB1</li>
 +
                    </ul>
 +
          </ul>
 
     </article>
 
     </article>
  
Line 2,003: Line 1,061:
 
     <article id="August-14" class="note-item">
 
     <article id="August-14" class="note-item">
 
         <h2>August 14</h2><hr>
 
         <h2>August 14</h2><hr>
       <p class="note-title">Transformation</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Cultivation</li>
 +
              <li class="list">Miniprep (Purify Plasmid)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Enterocin A + pTXB1</li>
                      <li class="list">OAIP-his 8/4 A<sup>r</sup></li>
+
                        <li class="list">Enterocin B + pTXB1</li>
                      <li class="list">Hv1a-his 7/29 A<sup>r</sup></li>
+
                        <li class="list">Enterocin 96 + pTXB1</li>
                      <li class="list">Sf1a-lectin-his 8/4 A<sup>r</sup></li>
+
                        <li class="list">Duracin TW49M + pTXB1</li>
                      <li class="list">OAIP-lectin-his 7/30  A<sup>r</sup></li>
+
                        <li class="list">Subtilosin + pTXB1</li>
                      <li class="list">Hv1a-lectin-his 8/4  A<sup>r</sup></li>
+
 
                 </ul>
 
                 </ul>
        </ul>
+
          </ul>
 
+
        <p class="note-title">Expression</p>
      <p class="note-title">PCR</p>
+
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Transformation (Transform correct plasmid into E. coli ER2566)to test different cultivation temperature</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>
 
         </ul>
 
     </article>
 
     </article>
Line 2,032: Line 1,082:
 
     <article id="August-15" class="note-item">
 
     <article id="August-15" class="note-item">
 
         <h2>August 15</h2><hr>
 
         <h2>August 15</h2><hr>
       <p class="note-title">PCR</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Purepose: Sequencing plasmid</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Enterocin A + pTXB1</li>
                      <li class="list">Hv1a-lectin-his 8/14 A<sup>r</sup> 1~6</li>
+
                        <li class="list">Enterocin B + pTXB1</li>
                      <li class="list">Sf1a- lectin-his 8/14 A<sup>r</sup> 1~6</li>
+
                        <li class="list">Enterocin 96 + pTXB1</li>
                      <li class="list">Sf1a-lectin-his 8/4 A<sup>r</sup> 1~6</li>
+
                        <li class="list">Duracin TW49M + pTXB1</li>
                      <li class="list">OAIP-lectin-his 7/30  A<sup>r</sup> 1~6</li>
+
                        <li class="list">Subtilosin + pTXB1</li>
                      <li class="list">OAIP- lectin-his 8/14  A<sup>r</sup> 1~6</li>
+
 
                 </ul>
 
                 </ul>
 
         </ul>
 
         </ul>
  
       <p class="note-title">Electrophoresis</p>
+
       <p class="note-title">Expression</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Purepose: Cultivation (Cultivation of E. coli ER2566 colonies for IPTG induction)</li>
              <li class="list">Sample</li>
+
               <li class="list">Purepose: IPTG Induction (To test the temperature of cultivation after induction)Condition: temp = 37, 30, 13.5°C</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>
 
         </ul>
 
     </article>
 
     </article>
Line 2,097: Line 1,104:
 
     <article id="August-16" class="note-item">
 
     <article id="August-16" class="note-item">
 
         <h2>August 16</h2><hr>
 
         <h2>August 16</h2><hr>
       <p class="note-title">Cultivation</p>
+
       <p class="note-title">Expression</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Purepose: SDS PAGE (To check the protein production after induction)Check different temperature</li>
              <li class="list">Sample</li>
+
               <li class="list">Purepose: IPTG Induction (To test the concentration of IPTG)Condition: concentration = 200, 400, 600, 800, 1000μM</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>
 
         </ul>
 
     </article>
 
     </article>
Line 2,154: Line 1,113:
 
     <article id="August-17" class="note-item">
 
     <article id="August-17" class="note-item">
 
         <h2>August 17</h2><hr>
 
         <h2>August 17</h2><hr>
           <p class="note-title">IPTG induction</p>
+
           <p class="note-title">Expression</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Purepose: SDS PAGE (To check the protein production after induction)Check different IPTG concentration</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
                <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
+
           </ul>
                      <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>
  
Line 2,192: Line 1,123:
 
         <h2>August 18</h2><hr>
 
         <h2>August 18</h2><hr>
 
       <p class="note-title">SDS-PAGE</p>
 
       <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>
  
 
     <article id="August-19" class="note-item">
 
     <article id="August-19" class="note-item">
 
         <h2>August 19</h2><hr>
 
         <h2>August 19</h2><hr>
         <p class="note-title">SDS-PAGE</p>
+
         <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Purepose: Digestion of PCR products (Including insert and backbone pET30a)</li>
               <li class="list">β-me make disulfide bond not folding.</li>
+
               <li class="list">Purepose: Ligation of digestion products (Ligase the DNA fragment into backbone pET30a)</li>
 
               <li class="list">Sample(8/17)</li>
 
               <li class="list">Sample(8/17)</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Enterocin A + pET30a</li>
                       <li class="list">Hv1a-lectin-his 500μM + β-me</li>
+
                       <li class="list">Enterocin B + pET30a</li>
                       <li class="list">Hv1a-lectin-his 500μM</li>
+
                       <li class="list">Enterocin 96 + pET30a</li>
                       <li class="list">Sf1a-lectin-his 0μM + β-me</li>
+
                       <li class="list">Bovicin HJ50 + pET30a</li>
                       <li class="list">Sf1a-lectin-his 500μM + β-me</li>
+
                       <li class="list">Duracin TW49M + pET30a</li>
                       <li class="list">Sf1a-lectin-his 500μM</li>
+
                       <li class="list">Lacticin Z + pET30a</li>
                      <li class="list">Rosetta-gami + β-me</li>
+
                       <li class="list">Leucocyclicin Q + pET30a</li>
                       <li class="list">OAIP-lectin-his 0μM + β-me</li>
+
                       <li class="list">Subtilosin + pET30a</li>
                       <li class="list">OAIP-lectin-his 500μM + β-me</li>
+
                      <li class="list">OAIP-lectin-his 500μM</li>
+
 
                 </ul>
 
                 </ul>
 
         </ul>
 
         </ul>
Line 2,234: Line 1,147:
 
     <article id="August-20" class="note-item">
 
     <article id="August-20" class="note-item">
 
         <h2>August 20</h2><hr>
 
         <h2>August 20</h2><hr>
       <p class="note-title">Sonication and SDS-PAGE</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Purepose: Transformation of ligation products (Transform into E. coli DH5α)</li>
 +
              <li class="list">Purepose: Taq PCR (PCR of ligation products to amplify insert gene)</li>
 +
              <li class="list">Purepose: Electrophoresis (To check PCR products)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Enterocin A + pET30a</li>
                       <li class="list">Hv1a-his A<sup>r</sup></li>
+
                       <li class="list">Enterocin B + pET30a</li>
                       <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
+
                       <li class="list">Enterocin 96 + pET30a</li>
                       <li class="list">Sf1a-lectin-his A<sup>r</sup></li>
+
                       <li class="list">Bovicin HJ50 + pET30a</li>
                       <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
+
                       <li class="list">Duracin TW49M + pET30a</li>
                       <li class="list">Hv1a-lectin-his A<sup>r</sup></li>
+
                       <li class="list">Lacticin Z + pET30a</li>
                       <li class="list">OAIP-lectin-his A<sup>r</sup></li>
+
                       <li class="list">Leucocyclicin Q + pET30a</li>
                       <li class="list">OAIP-lectin-his A<sup>r</sup></li>
+
                       <li class="list">Subtilosin + pET30a</li>
 
                 </ul>
 
                 </ul>
 
         </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>
 
 
     </article>
 
     </article>
  
 
     <article id="August-21" class="note-item">
 
     <article id="August-21" class="note-item">
 
         <h2>August 21</h2><hr>
 
         <h2>August 21</h2><hr>
       <p class="note-title">IPTG Induction and Sonication</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Purepose: Cultivation</li>
 +
              <li class="list">Purepose: Miniprep (Purify Plasmid)</li>
 
               <li class="list">Sample</li>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Enterocin A + pET30a</li>
                       <li class="list">Hv1a-lectin-his A<sup>r</sup> 8/20</li>
+
                      <li class="list">Enterocin B + pET30a</li>
                       <li class="list">Sf1a-his  A<sup>r</sup> 8/17</li>
+
                       <li class="list">Enterocin 96 + pET30a</li>
                       <li class="list">Sf1a-lectin-his A<sup>r</sup> 8/20</li>
+
                      <li class="list">Bovicin HJ50 + pET30a</li>
                       <li class="list">OAIP-his A<sup>r</sup> 8/17</li>
+
                       <li class="list">Duracin TW49M + pET30a</li>
                       <li class="list">OAIP-lectin-his A<sup>r</sup> 8/20</li>
+
                       <li class="list">Lacticin Z + pET30a</li>
 +
                       <li class="list">Leucocyclicin Q + pET30a</li>
 +
                       <li class="list">Subtilosin + pET30a</li>
 
                 </ul>
 
                 </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>
  
 
     <article id="August-22" class="note-item">
 
     <article id="August-22" class="note-item">
 
         <h2>August 22</h2><hr>
 
         <h2>August 22</h2><hr>
       <p class="note-title">Cultivation</p>
+
       <p class="note-title">Cloning</p>
 
           <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
           <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">Sequencing plasmid</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>
 
               <li class="list">Sample</li>
 
                 <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;">
 
                 <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">Enterocin A + pET30a</li>
                       <li class="list">BL21-Rosetta-gami 8/22</li>
+
                      <li class="list">Enterocin B + pET30a</li>
                       <li class="list">Sf1a-lectin-his 0μM 8/22</li>
+
                       <li class="list">Enterocin 96 + pET30a</li>
                       <li class="list">Sf1a-lectin-his 1000μM 8/22</li>
+
                      <li class="list">Bovicin HJ50 + pET30a</li>
 +
                       <li class="list">Duracin TW49M + pET30a</li>
 +
                      <li class="list">Lacticin Z + pET30a</li>
 +
                       <li class="list">Leucocyclicin Q + pET30a</li>
 +
                      <li class="list">Subtilosin + pET30a</li>
 
                 </ul>
 
                 </ul>
 
         </ul>
 
         </ul>

Revision as of 06:18, 27 September 2018

Navigation Bar

Wet Lab

July 1


Cloning

  • Received and resuspended of the IDT basic part
  • Sample
    • Enterocin A
    • Enterocin B
    • Enterocin 96
    • Bovicin HJ50
    • Duracin TW49M
    • Lacticin Z
    • Leucocyclicin Q
    • Subtilosin

July 2


None

July 3


Growth curve exp

  • Observe the growth population of Bacillis subtilis in different temperature Condition: 37°C, 32°C, 27°C

July 4


None

July 5


None

July 6


None

July 7


None

July 8


Growth curve exp

  • Observe the growth population of Bacillis subtilis in different pH Condition: pH = 4, 5, 6, 7, 8

July 9


None

July 10


None

July 11


None

July 12


None

July 13


None

July 14


None

July 15


Expression

  • repared competent cell E. coli ER2566

Growth curve exp

  • Observe the growth population of Bacillis subtilis in different salinity Condition: 0.17M, 0.25M, 0.5M, 0.75M, 1.0M
  • Sample

July 16


Cloning

  • Amplify the insert gene (Pfu PCR)
  • Electrophoresis (To check PCR products)
  • Sample
    • Lacticin Z + pTXB1
    • Bovincin HJ50+ pTXB1

July 17


Cloning

  • Digestion of PCR products (Including insert and backbone pTXB1)
  • Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
  • Sample
    • Lacticin Z + pTXB1
    • Bovincin HJ50+ pTXB1

July 18


Cloning

  • Transformation of ligation products (Transform into E. coli DH5α)
  • Taq PCR (PCR of ligation products to amplify insert gene)
  • Electrophoresis (To check PCR products)
  • Sample
    • Lacticin Z + pTXB1
    • Bovincin HJ50+ pTXB1

July 19


Cloning

  • Cultivation
  • Miniprep (Purify Plasmid)
  • Sample
    • Lacticin Z + pTXB1
    • Bovincin HJ50+ pTXB1

July 20


Cloning

  • PurposeL: Sequencing plasmid
  • Sample
    • Lacticin Z + pTXB1
    • Bovincin HJ50+ pTXB1

July 21


Cultivation

July 22


Cloning

  • Amplify the insert gene (Pfu PCR)
  • Electrophoresis (To check PCR products)
  • Digestion of PCR products (Including insert and backbone pTXB1)
  • Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
  • Sample
    • Leucocyclicin Q + pTXB1

July 23


Cloning

  • Transformation of ligation products (Transform into E. coli DH5α)
  • Taq PCR (PCR of ligation products to amplify insert gene)
  • Electrophoresis (To check PCR products)
  • Sample
    • Leucocyclicin Q + pTXB1

July 24


Cloning

  • Cultivation
  • Miniprep (Purify Plasmid)
  • Sample
    • Leucocyclicin Q + pTXB1

July 25


Cloning

  • Sequencing plasmid
  • Sample
    • Leucocyclicin Q + pTXB1

July 26


None

July 27


None

July 28


None

July 29


Expression

  • Transformation (Transform correct plasmid into E. coli ER2566)
    • Leucocyclicin Q + pTXB1

July 30


None

July 31


None

August 1


None

August 2


None

August 3


None

August 4


None

August 5


Expression

  • Purepose: Transformation (Transform correct plasmid into E. coli ER2566)
  • sample
    • Leucocyclicin Q + pTXB1

August 6


Expression

  • Purepose: Cultivation (Cultivation of E. coli ER2566 colonies for IPTG induction)
  • Purepose: IPTG Induction (To test the OD levels of E. coli ER2566 to induce)Condition: OD = 0.5, 1.3
  • sample
    • Leucocyclicin Q + pTXB1

August 7


Expression

  • Purepose: SDS PAGE (To check the protein production after induction)
  • sample
    • Leucocyclicin Q + pTXB1

August 8


None

August 9


None

August 10


None

August 11


Cloning

  • Purepose: Amplify the insert gene (Pfu PCR)
  • Purepose: Electrophoresis (To check PCR products)
  • sample
    • Enterocin A
    • Enterocin B
    • Enterocin 96
    • Duracin TW49M
    • Subtilosin

August 12


Cloning

  • Purepose: Digestion of PCR products (Including insert and backbone pTXB1)
  • Purepose: Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
  • Purepose: Transformation of ligation products (Transform into E. coli DH5α)
  • sample
    • Enterocin A + pTXB1
    • Enterocin B + pTXB1
    • Enterocin 96 + pTXB1
    • Duracin TW49M + pTXB1
    • Subtilosin + pTXB1

Growth curve

  • Purepose: Observe the growth of Bacillus subtilis in different temperature, pH, salinityCondition:
  • 1.Temp: 37°C , pH:7 , salinity: 0.17M
  • 2.Temp: 30°C , pH:9 , salinity: 0.5M
  • 3.Temp: 25°C , pH:5 , salinity: 0.25M

August 13


Cloning

  • Purepose: Taq PCR (PCR of ligation products to amplify insert gene)
  • Purepose: Electrophoresis (To check PCR products)
  • sample
    • Enterocin A + pTXB1
    • Enterocin B + pTXB1
    • Enterocin 96 + pTXB1
    • Duracin TW49M + pTXB1
    • Subtilosin + pTXB1

August 14


Cloning

  • Cultivation
  • Miniprep (Purify Plasmid)
  • Sample
    • Enterocin A + pTXB1
    • Enterocin B + pTXB1
    • Enterocin 96 + pTXB1
    • Duracin TW49M + pTXB1
    • Subtilosin + pTXB1

Expression

  • Transformation (Transform correct plasmid into E. coli ER2566)to test different cultivation temperature

August 15


Cloning

  • Purepose: Sequencing plasmid
  • Sample
    • Enterocin A + pTXB1
    • Enterocin B + pTXB1
    • Enterocin 96 + pTXB1
    • Duracin TW49M + pTXB1
    • Subtilosin + pTXB1

Expression

  • Purepose: Cultivation (Cultivation of E. coli ER2566 colonies for IPTG induction)
  • Purepose: IPTG Induction (To test the temperature of cultivation after induction)Condition: temp = 37, 30, 13.5°C

August 16


Expression

  • Purepose: SDS PAGE (To check the protein production after induction)Check different temperature
  • Purepose: IPTG Induction (To test the concentration of IPTG)Condition: concentration = 200, 400, 600, 800, 1000μM

August 17


Expression

  • Purepose: SDS PAGE (To check the protein production after induction)Check different IPTG concentration
  • Sample

August 18


SDS-PAGE

August 19


Cloning

  • Purepose: Digestion of PCR products (Including insert and backbone pET30a)
  • Purepose: Ligation of digestion products (Ligase the DNA fragment into backbone pET30a)
  • Sample(8/17)
    • Enterocin A + pET30a
    • Enterocin B + pET30a
    • Enterocin 96 + pET30a
    • Bovicin HJ50 + pET30a
    • Duracin TW49M + pET30a
    • Lacticin Z + pET30a
    • Leucocyclicin Q + pET30a
    • Subtilosin + pET30a

August 20


Cloning

  • Purepose: Transformation of ligation products (Transform into E. coli DH5α)
  • Purepose: Taq PCR (PCR of ligation products to amplify insert gene)
  • Purepose: Electrophoresis (To check PCR products)
  • Sample
    • Enterocin A + pET30a
    • Enterocin B + pET30a
    • Enterocin 96 + pET30a
    • Bovicin HJ50 + pET30a
    • Duracin TW49M + pET30a
    • Lacticin Z + pET30a
    • Leucocyclicin Q + pET30a
    • Subtilosin + pET30a

August 21


Cloning

  • Purepose: Cultivation
  • Purepose: Miniprep (Purify Plasmid)
  • Sample
    • Enterocin A + pET30a
    • Enterocin B + pET30a
    • Enterocin 96 + pET30a
    • Bovicin HJ50 + pET30a
    • Duracin TW49M + pET30a
    • Lacticin Z + pET30a
    • Leucocyclicin Q + pET30a
    • Subtilosin + pET30a

August 22


Cloning

  • Sequencing plasmid
  • Sample
    • Enterocin A + pET30a
    • Enterocin B + pET30a
    • Enterocin 96 + pET30a
    • Bovicin HJ50 + pET30a
    • Duracin TW49M + pET30a
    • Lacticin Z + pET30a
    • Leucocyclicin Q + pET30a
    • Subtilosin + pET30a

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