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Team:Yale/Our Project/Notebook/Week 4
From 2010.igem.org
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| - | Monday | + | Monday 6/28--PCR amplification of thiosulfate reductase operon phsABC |
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| - | + | <b> pSB74 transformants<b> <br/> | |
| + | Extensive colony growth was seen on plated transformations of pSB74 into DH5alpha and LE392 from <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_3"> 6/25 </a> but none was visible on BL21 plate. Continued failure of BL21 transformation suggests that the homemade competent BL21 cells being used are somehow faulty. <br/> | ||
| + | <h4> PCR amplification of phsABC </h4> | ||
| + | <ul> | ||
| + | <li> Prior to PCR, took aliquots of all primer samples and diluted them first to 100 uM as detailed below by the chart and then again to 10 uM </li> | ||
| + | <table> | ||
| + | <tr> | ||
| + | <td>Primer</td> <td> phsABC_F </td> <td> phsABC_R </td> <td> phsAB_R </td> <td> phsC_F </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td> Initial Amount </td> <td>38.56 nm</td> <td>46.58 nm</td><td>29.03 nm</td><td>21.94 nm</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td> Volume of water </td> <td>385.6 uL</td> <td>465.8 uL</td><td>290.3 uL</td><td>219.4 uL</td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | <li> Similarly, diluted the DNA sample to 10 ng DNA/uL. Started with 0.5 uL of miniprepped pSB74 (sample a) at 67.3 ng/uL and added 2.865 uL of water to get the desired concentration.</li> | ||
| + | <t | ||
| + | </ul> | ||
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Revision as of 09:12, 27 October 2010
our project
lab notebook: week 4 (6/28 -7/4)
- Monday 6/28--PCR amplification of thiosulfate reductase operon phsABC See more/less
- Tuesday short content See more/less
- Wednesday short content See more/less
- Thursday short content See more/less
- Fridays short content See more/less
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