Team:NYMU-Taipei/notebook/labnotes

From 2014.igem.org

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<h1>lab note of circuit in the cohesion part</h1>
<h1>lab note of circuit in the cohesion part</h1>
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<p>part/gene transformation and test</p>
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<h2>Working Circuit Assembling</h2>
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<p>circuit contruction</p>
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<p><b>1. Check IGEM standard part-K523013(trunk).B0015</b></p>
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<p>part function test</p>
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    <ol>
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<p>under construction</p>
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      <li>IGEM kit transformation with DH5 competent cell</li>
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<h2>experiment1 date: 0707~0712</h2>
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      <li>colony PCR check: pick up colony of transformation and do the PCR to check part length</li>
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<p>under construction</p>
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      <li>plasmid PCR check: extract the plasmid of the correct length of part after checking colony PCR check, and use plasmid PCR to check length again</li>
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<p>under construction</p>
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      <li>Digestion check: use enzyme to cut plasmid of correct length of PCR, to check restriction site</li>
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<p>under construction</p>
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      <li>Second plate and liquid culture save: save bacteria containing correct plasmid in liquid culture and second plate</li>
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<p>under construction</p>
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      <img src=''>
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<h2>experiment1 date: 0707~0712</h2>
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    </ol>
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<p>under construction</p>
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<p><b>2. Get S. mutans competent signal peptide (CSP16) part</b></p>
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<p>under construction</p>
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    <ol>
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<p>under construction</p>
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      <li>S. mutans genome extraction </li>
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<h2>experiment1 date: 0707~0712</h2>
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      <li>Tag PCR to get CSP16 linear DNA and check length through electrophoresis: we use Tag PCR to test the condition of the most suitable condition of PCR</li>
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<p>under construction</p>
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      <li>KOD PCR to get CSP16 linear DNA, check length through electrophoresis, and gel extraction to get CSP16 part: in the condition tested during Tag PCR, we use designed primer and KOD PCR to amplify and copy CSP16 linear DNA , and run electrophoresis and gel extraction to get CSP16 part</li>
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<p>under construction</p>
+
      <li>Digestion check: use enzyme to cut plasmid of correct length of PCR, to check restriction site</li>
 +
      <li>Second plate and liquid culture save: save bacteria containing correct plasmid in liquid culture and second plate</li>
 +
    </ol>
 +
<p><b>3. K523013(trunk) and CSP16 assembling</b></p>
 +
    <ol>
 +
      <li>We use Tag PCR with designed primer to check suitable PCR condition</li>
 +
      <li>We conducted KOD PCR with designed primer to get K523013 assembled with CSP16 linear DNA and check length through electrophoresis, and gel extraction to get CSP16 plus K523013(trunk) part: in the condition tested during Tag PCR, we use designed primer and KOD PCR to assemble CSP16 linear DNA and K523013(trunk) , and run electrophoresis and gel extraction to get CSP16 assembled with K523013(trunk) part</li>
 +
      <img src=''>
 +
      <img src=''>
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    </ol>
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<p><b>4. K523013 assembled with CSP16 and BBa-B0015 assembling</b></p>
 +
    <ol>
 +
      <li>K523013 assembled with CSP16 digestion: we use ECOR1 and SPET1 enzyme to digest K523013 assembledCSP16</li>
 +
      <li>BBa-B0015 digestion: we use ECOR1 and XBAL1 enzyme to digest BBa-B0015</li>
 +
      <li>K523013 assembled with CSP16and BBa-B0015 ligation: we use T4 ligase to ligase K523013 assembled with CSP16and BBa-B0015</li>
 +
      <li>transformation with DH5ɚ competent cell</li>
 +
      <li>After plasmid extraction of the ciucuit ,it ran PCR and electrophoresis to check length of circuit</li>
 +
      <li>digestion to check circuit length and restriction site </li>
 +
      <li>plasmid sequencing of the final circuit</li>
 +
      <img src=''>
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    </ol>
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<h2>Testing Circuit Assembling</h2>
         </div>
         </div>
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Revision as of 17:38, 29 September 2014

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Lab notes
introduction
control
communication
cohesion
completion
function test

introduction :P introduction :P introduction :P introduction :P

introduction of labnote

樹狀圖 timeline 順序圖

under construction

under construction

under construction

under construction

under construction

under construction

under construction

under construction

under construction

under construction

under construction

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under construction

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