Difference between revisions of "Team:UofGuelph/Contributions"
| Line 117: | Line 117: | ||
<th>Component</th> | <th>Component</th> | ||
<th>Volume (µL)</th> | <th>Volume (µL)</th> | ||
| − | + | </tr> | |
| − | + | ||
<tr> | <tr> | ||
<td>H<sub>2</sub>O</td> | <td>H<sub>2</sub>O</td> | ||
<td>20.4</td> | <td>20.4</td> | ||
| − | + | </tr> | |
| − | + | ||
<tr> | <tr> | ||
<td>Phusion Buffer</td> | <td>Phusion Buffer</td> | ||
<td>8.0</td> | <td>8.0</td> | ||
| − | + | </tr> | |
| − | + | ||
<tr> | <tr> | ||
<td>dNTPs</td> | <td>dNTPs</td> | ||
<td>0.8</td> | <td>0.8</td> | ||
| − | + | </tr> | |
| − | + | ||
<tr> | <tr> | ||
<td>VF2</td> | <td>VF2</td> | ||
<td>0.2</td> | <td>0.2</td> | ||
| − | + | </tr> | |
| − | + | ||
<tr> | <tr> | ||
<td>VR</td> | <td>VR</td> | ||
<td>0.2</td> | <td>0.2</td> | ||
| − | + | </tr> | |
| − | + | ||
<tr> | <tr> | ||
<td>BBa_DNA</td> | <td>BBa_DNA</td> | ||
<td>10</td> | <td>10</td> | ||
| − | + | </tr> | |
| − | + | ||
<tr> | <tr> | ||
<td>Phusion</td> | <td>Phusion</td> | ||
<td>0.4</td> | <td>0.4</td> | ||
| − | + | </tr> | |
| − | + | ||
<tr> | <tr> | ||
<td><b>Total</b></td> | <td><b>Total</b></td> | ||
<td><b>40</b></td> | <td><b>40</b></td> | ||
| − | + | </tr> | |
| − | + | ||
</table> | </table> | ||
| + | <p class="descP"> | ||
| + | PCR Cycling Parameters: | ||
| + | </p> | ||
| + | |||
| + | |||
| + | <table style="width:20%" align="centre"> | ||
| + | <tr> | ||
| + | <th>Step</th> | ||
| + | <th>Temperature (°C)</th> | ||
| + | <th>Time (minutes)</th> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Initial Denaturation</td> | ||
| + | <td>98</td> | ||
| + | <td>2</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Denaturation</td> | ||
| + | <td>98</td> | ||
| + | <td>1</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Annealing</td> | ||
| + | <td>60</td> | ||
| + | <td>1</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Extension</td> | ||
| + | <td>72</td> | ||
| + | <td>0.5</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>Final Extension</td> | ||
| + | <td>72</td> | ||
| + | <td>10</td> | ||
| + | </tr> | ||
| + | |||
| + | </table> | ||
Revision as of 03:13, 18 October 2018
Registry Contributions
2018 Registry Submissions
Sequencing of BBa_K747047
Background
iGEM Guelph's 2018 contribution to the registry was the sequence analysis of a BioBrick located in the 2018 Distribution Kit.
BioBrick K747047 was located in Plate 2, Well 12O. Our team chose to sequence this part as the Distribution Kit listed it as a Bad Sequence and for our team’s first characterization of a Registry Part, we thought that this part was a good candidate. Sequencing preparation was performed by our team, while the sequencing was performed at the Laboratory Services at the University of Guelph.
Methods
A PCR was set up according to the following recipe:
| Component | Volume (µL) |
|---|---|
| H2O | 20.4 |
| Phusion Buffer | 8.0 |
| dNTPs | 0.8 |
| VF2 | 0.2 |
| VR | 0.2 |
| BBa_DNA | 10 |
| Phusion | 0.4 |
| Total | 40 |
PCR Cycling Parameters:
| Step | Temperature (°C) | Time (minutes) |
|---|---|---|
| Initial Denaturation | 98 | 2 |
| Denaturation | 98 | 1 |
| Annealing | 60 | 1 |
| Extension | 72 | 0.5 |
| Final Extension | 72 | 10 |
