Team:NYMU-Taipei/notebook/labnotes

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       <h>Lab notes</h>
       <h>Lab notes</h>
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      <div id='tabframe'>
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<img src='/wiki/images/7/7b/NYMU14_fNotebookSmall.png'>
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        <div class='note-tab' id='nt1'>introduction</div>
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        <div class='note-tab' id='nt2'>control</div>
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        <div class='note-tab' id='nt3'>communication</div>
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        <div class='note-tab' id='nt4'>cohesion</div>
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        <div class='note-tab' id='nt5'>completion</div>
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        <div class='note-tab' id='nt6'>function test</div>
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      </div>
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       <div id='labnote'>
       <div id='labnote'>
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         <div class='note-tab'><p>labnote 1</p></div>
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         <div class='note-content notenow' id='nt1-cont'>
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         <div class='note-tab'><p>labnote 2</p></div>
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<h1>introduction :P introduction :P introduction :P introduction :P </h1>
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         <div class='note-tab'><p>labnote 3</p></div>
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<p>introduction of labnote</p>
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         <div class='note-tab'><p>labnote 4</p></div>
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<p>樹狀圖 timeline 順序圖</p>
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<p>under construction</p>
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        </div>
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         <div class='note-content' id='nt2-cont' style='display:none;'>
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<h1>lab note of circuit in the control part</h1>
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<p>part/gene transformation and test</p>
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<p>circuit contruction</p>
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<p>part function test</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<h2>experiment1 date: 0707~0712</h2>
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<h2>experiment1 date: 0707~0712</h2>
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        </div>
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         <div class='note-content' id='nt3-cont' style='display:none;'>
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<h1>lab note of circuit in the communication part</h1>
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<p>part/gene transformation and test</p>
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<p>circuit contruction</p>
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<p>part function test</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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<p>under construction</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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        </div>
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         <div class='note-content' id='nt4-cont' style='display:none;'>
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<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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<p>circuit contruction</p>
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<p>part function test</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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<p>under construction</p>
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<p>under construction</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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<p>under construction</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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<p>under construction</p>
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        </div>
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        <div class='note-content' id='nt5-cont' style='display:none;'>
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<h1>lab note of circuit in the completion part</h1>
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<h2>Parts from iGEM took kit</h2>
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<h3>Out line</h3>
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<p>For each part to be used in testing circuit:<br>BBa_J23100 , BBa_E1010, BBa_B0015
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</p>
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<img src=''><p>circuit 圖</p>
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<p>After transforming these plasmids into our competent cell, DH5a, we would take 3 steps to examine if we got the right product:</p>
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<ol>
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<li>Colony PCR and electrophoresis: Directly picking up colonies on designated antibiotic plate and do colony PCR with iGEM provided primers (VF2&VR), and then do electrophoresis to check the length of plasmid to confirm if the kit had been correctly transformed into E. coli.</li>
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<li>Plasmid PCR and electrophoresis: Colony PCR may not end up copying the right place, for there is extra DNA in the cell that is not the one of our interest. Therefore, we also check the length of the plasmid we which we transformed.</li>
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<li>Digestion and electrophoresis: After checking the length, we use EcoR1/Spe1 and Xba1/Pst1, and electrophoresis, to see if our constructed plasmid can be successfully digested.</li>
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</ol>
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<h3>Process</h3>
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<p>We encounter several difficulties during the process:</p>
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<ol>
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<li>In the transformation step, we couldn’t get transformation with the right PCR band at first. Even thought they do grow on the antibiotic plate, electrophoresis result doesn’t show any band. Therefore, we tried different ways trying to solve these problems.<br>First, we thought it was PCR problem, therefore, we tried colony PCR. However, it came out that it wasn’t the case.So we improve our transformation protocol. That is, after 1.5 hour of recovery in 37C incubator in 200ml LB, we centrifuge in 3.4 rpm for 20 seconds, and discard 150ml supernatant.<br>At the end, we result in putting concentrated transformed E.coli into our antibiotic plate. This way, we sequentially transformed several kits successfully.</li>
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<li>In the checking step, we’ve have contaminated electrophoresis result after colony PCR. That is, we got band in the negative well. To find out the ingredient causing contamination, we do several negative control changing different packing of ingredients. And find the contamination of our 10X DreamTaq Buffer.</li>
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</ol>
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<h3>Result</h3>
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<img src='/wiki/images/4/43/NYMU14_note_4c_r1.PNG' style='width:430px;'>
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<img src='/wiki/images/f/ff/NYMU14_note_4c_r2.PNG' style='width:430px;'>
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<img src='/wiki/images/a/a3/NYMU14_note_4c_r3.PNG'>
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<img src='/wiki/images/c/cd/NYMU14_note_4c_r4.PNG'>
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<img src='/wiki/images/e/eb/NYMU14_note_4c_r5.PNG'>
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        </div>
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        <div class='note-content' id='nt6-cont' style='display:none;'>
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<h1>lab note of function test in phage</h1>
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<p>part/gene transformation and test</p>
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<p>circuit contruction</p>
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<p>part function test</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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<p>under construction</p>
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<p>under construction</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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<p>under construction</p>
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<p>under construction</p>
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<h2>experiment1 date: 0707~0712</h2>
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<p>under construction</p>
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<p>under construction</p>
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        </div>
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       </div>
       </div>
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      <embed src='/wiki/images/8/87/NYMU14_Labnote_Target1.pdf' class='note-pdf' style='display:block;'>
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      <embed src='/wiki/images/c/c6/NYMU14_Labnote_Target2.pdf' class='note-pdf' style='display:none;'>
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      <embed src='/wiki/images/e/eb/NYMU14_Labnote_Target3.pdf' class='note-pdf' style='display:none;'>
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      <embed src='/wiki/images/9/93/NYMU14_labnote_inhibitor1.pdf' class='note-pdf' style='display:none;'>
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      <embed src='' class='note-pdf' style='display:none;'>
   </div>
   </div>
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Latest revision as of 22:04, 17 October 2014

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Lab notes

labnote 1

labnote 2

labnote 3

labnote 4

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