Difference between revisions of "Team:NYU Shanghai/Protocols"
(11 intermediate revisions by the same user not shown) | |||
Line 478: | Line 478: | ||
<div id="chromoText" style="display:none"> | <div id="chromoText" style="display:none"> | ||
<p> | <p> | ||
− | <h6><font color="#d66">Building our Construct</font></h6> | + | <h6><font color="#d66">Building our Construct: from biobrick parts in the kit</font></h6> |
− | <p>Note: If using construct with | + | <p>Note: If using construct with pBAD promoter, DO NOT USE SOC MEDIA. Glucose inhibits the uptake of arabinose, and will inhibit promoter induction. |
+ | <br>Note: We should have used PCR to amplify linearized backbone. | ||
+ | <br>Note: Always use gel electrophoresis to check digest results.</p> | ||
<br><img src="https://static.igem.org/mediawiki/2015/d/df/NYU_Shanghai_Chromo_Procedure.png" width="800"> | <br><img src="https://static.igem.org/mediawiki/2015/d/df/NYU_Shanghai_Chromo_Procedure.png" width="800"> | ||
+ | <br><br><br> | ||
+ | <h6><font color="#d66">Building our Construct: from IDT gBlocks</font></h6> | ||
+ | <p>Note: We recommend adding a reporter gene to the construct.</p> | ||
+ | <br><img src="https://static.igem.org/mediawiki/2015/7/71/NYU_Shanghai_IDTprocedure.png" width="550"> | ||
<br><br><br> | <br><br><br> | ||
<h6><font color="#d66">Expressing XJTLU's Construct</font></h6> | <h6><font color="#d66">Expressing XJTLU's Construct</font></h6> | ||
Line 926: | Line 932: | ||
<td>Nuclease-free water</td> | <td>Nuclease-free water</td> | ||
<td>10μl or none</td> | <td>10μl or none</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | <br> | ||
+ | <table> | ||
+ | <tr> | ||
+ | <td><font color="#d66">Initial Denaturation</font></td> | ||
+ | <td>98C</td> | ||
+ | <td>30s</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><font color="#d66">25 cycles</font></td> | ||
+ | <td>98C</td> | ||
+ | <td>15s</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Annealing temp 1</td> | ||
+ | <td>59.5C</td> | ||
+ | <td>30s</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Annealing temp 2</td> | ||
+ | <td>56.3C</td> | ||
+ | <td>30s</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Annealing temp 3</td> | ||
+ | <td>53.7C</td> | ||
+ | <td>30s</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Extension</td> | ||
+ | <td>72C</td> | ||
+ | <td>60s</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><font color="#d66">Final Extension</font></td> | ||
+ | <td>72C</td> | ||
+ | <td>2m</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><font color="#d66">Hold</font></td> | ||
+ | <td>4C</td> | ||
<td></td> | <td></td> | ||
</tr> | </tr> |
Latest revision as of 16:56, 18 September 2015