|
|
| Line 10: |
Line 10: |
| | | | |
| | <div class="column full_size"> | | <div class="column full_size"> |
| − | <hr>
| + | <div align="center> |
| − | <div align="center"> | + | <a href="https://2016.igem.org/Team:Stony_Brook/Notebook/Week1" <h1> Week 1: 6/28-7/1 </h1> </a> |
| − | <h1> Week 1 (6/27 - 7/2) </h1> | + | |
| − | <h3 style="color:#B61C1D;"> 6/27 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> PCR of 3xHA-TDGF1 construct. 12 cycles </li>
| + | |
| − | <li> Gel of PCR Product → PCR was a failure. Band was 300bp. Should have been 750bp</li>
| + | |
| − | <li> Second PCR of 3xHA-TDGF1 construct. 25 cycles instead of 12 → left overnight </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h5> Interlab Study </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Measured LUDOX and H<sub>2</sub>O for plate reader </li>
| + | |
| − | <li> Transformation of positive and negative controls, test devices 1-3 </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;"> 6/28</h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Ran gel on PCR of yesterday's construct → Gel worked </li>
| + | |
| − | <li> Gel purification of PCR Product → low concentration of DNA found during nanodrop</li>
| + | |
| − | <li> PCR the construct using a revised method </li>
| + | |
| − | <li> 2:54pm → ran the gel on the PCR product </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h5> Interlab Study </h5>
| + | |
| − | <ul>
| + | |
| − | <li align> Heat and "SB" streaking seems not to have worked</li>
| + | |
| − | <li align> No visible red colonies on plate </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;"> 6/29 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> PCR Not working well → keep trying new methods with different annealing temperatures </li>
| + | |
| − | <li> Tried 65 and 67 degrees C</li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h5> Interlab Study </h5>
| + | |
| − | <ul>
| + | |
| − | <li> FITC started </li>
| + | |
| − | <li> Cutting re-inoculated </li>
| + | |
| − | <li> Ordered new LUDOX </li>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h5> Other </h5>
| + | |
| − | <ul>
| + | |
| − | <li> College of Arts and Sciences Pre College Summer Institute students stopped by lab </li>
| + | |
| − | <li> Spoke to them about information on synthetic biology and iGEM </li>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;"> 6/30 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> PCR worked → Test 5 → changed annealing temperature to 66 degrees C + increased the denaturing time </li>
| + | |
| − | <li> Nanodropped DNA</li>
| + | |
| − | <ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;"> 7/1 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Digestion of TDGF-1 construct and YFP352GAP vector with two restriction enzymes </li>
| + | |
| − | <li> Will ligate later </li>
| + | |
| − | </ul>
| + | |
| − | <table border=1>
| + | |
| − | <tr>
| + | |
| − | <th></th>
| + | |
| − | <th>TDGF-1 Construct (Phire PCR 128.2 ng/mL)</th>
| + | |
| − | <th>YEP352GAP Vector (343.2 ng/mL)</th>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Total Reaction Volume</td>
| + | |
| − | <td>50 uL</td>
| + | |
| − | <td>50 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Nuclease Free Water</td>
| + | |
| − | <td>27.4 uL</td>
| + | |
| − | <td>37.2 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>10x Cutsmart Buffer</td>
| + | |
| − | <td>5 uL</td>
| + | |
| − | <td>5uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>xho1</td>
| + | |
| − | <td>1 uL</td>
| + | |
| − | <td>1 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Pvu II</td>
| + | |
| − | <td>1 uL</td>
| + | |
| − | <Td>1 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>DNA</td>
| + | |
| − | <td>15.6 uL</td>
| + | |
| − | <td>5.8 uL</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;">7/2 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Running yesterday's digestion on gel </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h1> Week 2 (7/5-7/8) </h1> | + | |
| − | <h3 style="color:#B61C1D;">7/5 </h3>
| + | |
| − | </dv>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Ran the PCR'ed construct on a gel with Phire Polymerase → It didn't work
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;">7/6 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Ran the construct with PCR on gel → it worked </li>
| + | |
| − | <li> Tasnia stabbed our gel </li>
| + | |
| − | <li> Interlab study continues </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;">7/7 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Nanodrops were pretty low (84-120 ng/nl) </li>
| + | |
| − | <li> 11:46am PCR'ed the highest nanodrop result in tube #1 → PCR successful </li>
| + | |
| − | <li> 2:33pm → attempt to digest with remaining non-PCR product → removed from incubator at 5:00pm </li>
| + | |
| − | <li> Made a gel → ran PCR </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h1> Week 3 (7/11-7//15) </h1>
| + | |
| − | <h3 style="color:#B61C1D;"> 7/11 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Ran PCR on the construct (5x) → gel did not work </li>
| + | |
| − | <li> PCR'ed 84.2 ng/ml construct </li>
| + | |
| − | <li> 5 replicates made using Phire </li>
| + | |
| − | <li>Thermocycler Program</li>
| + | |
| − | </ul>
| + | |
| − | <br>
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <th colspan=3 align=center>Thermocycler Program</th>
| + | |
| − | <tr>
| + | |
| − | <th>Step</th>
| + | |
| − | <th>Temperature (Degrees C)</th>
| + | |
| − | <th>Duration (seconds)</th>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>1</td>
| + | |
| − | <td>98</td>
| + | |
| − | <td>30</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>2</td>
| + | |
| − | <td>98</td>
| + | |
| − | <td>20</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>3</td>
| + | |
| − | <td>66</td>
| + | |
| − | <td>5</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>4</td>
| + | |
| − | <td>72</td>
| + | |
| − | <td>30</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>5</td>
| + | |
| − | <td>72</td>
| + | |
| − | <td>60</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>6</td>
| + | |
| − | <td>4</td>
| + | |
| − | <td>Hold</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td colspan=3>After step 4, return to step 2. Repeat steps 2-4 thirty-five times before continuing to step 5 and on.</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | | + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Water</td>
| + | |
| − | <td>19.5 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Phire Polymerase</td>
| + | |
| − | <td>25 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Template</td>
| + | |
| − | <td>0.5 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Forward Primer</td>
| + | |
| − | <td>2.5 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Reverse Primer</td>
| + | |
| − | <td>2.5 uL</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | | + | |
| − | <ul>
| + | |
| − | <li> Restreaking of YEP352GAP transformed cells for miniprep</li>
| + | |
| − | <li> Restreaking placed in incubator at 12am</li>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Stuff & Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;"> 7/12 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Made gel for electrophoresis of PCR construct → it didn't work </li>
| + | |
| − | <li>Thermocycler Program: Same as PCR from 7/11</li>
| + | |
| − | </ul>
| + | |
| − | <br>
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <td>Q5 Rx Buffer</td>
| + | |
| − | <td>10 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>10nM dNTPs</td>
| + | |
| − | <td>1 uL</td>
| + | |
| − | </tr> | + | |
| − | <tr>
| + | |
| − | <td>10 uM Forward Primer</td>
| + | |
| − | <td>2.5 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>10 uM Reverse Primer</td>
| + | |
| − | <td>2.5 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Template (Q5 PCR Pw.1 for 6/30/16)</td>
| + | |
| − | <td>0.5 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Q5 Polymerase</td>
| + | |
| − | <td>2.5 uL</td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td>Water</td>
| + | |
| − | <td>33 uL</td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | <ul>
| + | |
| − | <li> LB liquid culture (3ml with 3ul of ampicillin) </li>
| + | |
| − | <li> Loaded 10ul Ladder DNA. </li>
| + | |
| − | <li> Loaded 1ul dye + 5ul PCR product </li>
| + | |
| − | <li> Ran gel at 115V </li>
| + | |
| − | <li> Checked gel ladder → faint, no band for the PCR product </li>
| + | |
| − | <li> Made new gel, loaded with the same amount of reagents as before → ran at 90V → No ladder band </li>
| + | |
| − | <li> Ran it again at 115V → visible ladder, no PCR band </li>
| + | |
| − | <li> Preparing a gel to run ladder and construct that previously showed a strong band → used this to diagnose a problem with gel setup. Ran TDGF1 152.9 Phire Child. Lane 1 is ladder, Lane 2 is that construct (10ul) with 2ul of dye </li>
| + | |
| − | <li> Setup new PCR using the 6/30/16 PCR construct </li>
| + | |
| − | </ul>
| + | |
| − | <ol>
| + | |
| − | <li> 98°C → 30sec </li>
| + | |
| − | <li> 98°C → 20sec </li>
| + | |
| − | <li> 66°C → 5sec </li>
| + | |
| − | <li> 72°C → 30sec </li>
| + | |
| − | <li> 72°C → 1min </li>
| + | |
| − | <li> 4°C → hold </li>
| + | |
| − | </ol>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Stuff & Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;">7/13 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Ran 5ul of PCR with ladder, cancer construct and vaccine construct → to diagnose problem with gel </li>
| + | |
| − | <li> Miniprep of Dean vector (3ml LB culture) </li>
| + | |
| − | <li> Purification of PCR product → nanodropped 16.4ug/ul </li>
| + | |
| − | <li> Nanodrop of: </li>
| + | |
| − | </ul>
| + | |
| − | <ol>
| + | |
| − | <li> Yellow 7/13 construct NEB purified → 16.4ng/ml</li>
| + | |
| − | <li> Blue gel purification → 48.7ng/ul</li>
| + | |
| − | <li> Pink PCR purification 7/13 → 102.2ng/ul</li>
| + | |
| − | <li>7/13 -RK Vector 1 → 48.2ng/ul</li>
| + | |
| − | <li> 7/13 -RK Vector 2 → 96.3ng/ul</li>
| + | |
| − | </ol>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Things </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;"> 7/14</h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer group </h5>
| + | |
| − | <ul>
| + | |
| − | <li>Things</li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine Group </h5>
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <th> Contents of well </th>
| + | |
| − | <th> Amount (ul)</th>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td> Ladder </td>
| + | |
| − | <td> 10 </td>
| + | |
| − | </tr>
| + | |
| − | <td> AD </td>
| + | |
| − | <td> 5 </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td> AO </td>
| + | |
| − | <td> 5 </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td> AM </td>
| + | |
| − | <td> 5 </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td> AN </td>
| + | |
| − | <td> 5 </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td> Gal-Sunil </td>
| + | |
| − | <td> Added 1ul loading dye </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td> Con-Sunil </td>
| + | |
| − | <td> 5 </td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | <ul>
| + | |
| − | <li> Brian and Sunil's Gal promoters worked but both constructs failed</li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h5> Biobrick ideas </h5>
| + | |
| − | <ol>
| + | |
| − | <li> Smelly yeast → optimized from e. coli </li>
| + | |
| − | <li> Twist on smelly e. coli on e. coli → regulated with quorum sensing</li>
| + | |
| − | <li> OmpA/EnvZ recognize osmolarity</li>
| + | |
| − | <li>Genetic Circuit</li>
| + | |
| − | <li> Shine yellow light → Banana smell. Shine green light → wintergreen </li>
| + | |
| − | <li> TetR operator system genetic circuit </li>
| + | |
| − | <li> Have cell die at certain wavelength → killswitch </li>
| + | |
| − | </ol>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <div align="center">
| + | |
| − | <h3 style="color:#B61C1D;"> 7/15 </h3>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Cancer group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Ran a gel on PCR products → </li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | <div class="column half_size">
| + | |
| − | <h5> Vaccine group </h5>
| + | |
| − | <ul>
| + | |
| − | <li> Jon and Sunil retrying construct </li>
| + | |
| − | <li> Jon used phire at 50°C</li>
| + | |
| − | <li> Sunil used Q5. Extension at 72°C for 30 seconds </li>
| + | |
| − | </ul>
| + | |
| − | <table>
| + | |
| − | <tr>
| + | |
| − | <th> Polymerase </th>
| + | |
| − | <th> Primers (ul) </th>
| + | |
| − | <th> Template (ul) </th>
| + | |
| − | <th> H<sub>2</sub>O (ul)</th>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td> Phire (25ul) </td>
| + | |
| − | <td> 2.5 Reverse Construct 2.5 Forward Stitch </td>
| + | |
| − | <td> 1.5 </td>
| + | |
| − | <td>19.5 </td>
| + | |
| − | </tr>
| + | |
| − | <tr>
| + | |
| − | <td> Q5 (25ul) </td>
| + | |
| − | <td> 2.5 Reverse Construct 2.5 Forward Stitch </td>
| + | |
| − | <td> 1.5 </td>
| + | |
| − | <td> 19.5 </td>
| + | |
| − | </tr>
| + | |
| − | </table>
| + | |
| − | </div>
| + | |
| − | | + | |
| − | | + | |
| − | <div class="column full_size">
| + | |
| − | <hr>
| + | |
| − | </div>
| + | |
| − | </html>
| + | |
