|
|
(18 intermediate revisions by 2 users not shown) |
Line 8: |
Line 8: |
| <div id="main"> | | <div id="main"> |
| | | |
− | <h5>Experiments & Protocols</h5>
| |
− | <div style="height:50px"></div>
| |
− |
| |
− | <!--
| |
− | <p>Describe the experiments, research and protocols you used in your iGEM project.</p>
| |
− |
| |
− | <ul>
| |
− | <li> Protocols </li>
| |
− | <li> Experiments </li>
| |
− | <li>Documentation of the development of your project </li>
| |
− | </ul>
| |
− | -->
| |
− |
| |
− | <div id="transformation"><h2>Transformation protocol </h2>
| |
− | <ol>
| |
− | <li>(If using part from the distribution: resuspend the DNA in 10 ul of MiliQ water, making sure that it turns red. Wait 10 minutes before adding the DNA to cells)</li>
| |
− | <li>Put a tube of NEB DH 5 alpha <i>E. coli</i> cells on ice and wait until they thaw completely. Divide the cells into 50 ul aliquotes. </li>
| |
− | <li>Add 1 ul of plasmid DNA to 50 ul of cells. </li>
| |
− | <li>Mix by carefully flicking the tube. Do not vortex or pipette in and out! </li>
| |
− | <li>Place the mixture on ice for 30 minutes. </li>
| |
− | <li>Heat shock the cells at 42 °C for 30 seconds and immediately put on back on ice.</li>
| |
− | <li>Keep cells on ice for next 5 minutes. Do not mix. </li>
| |
− | <li>Pipette 950 ul of SOC (or LB) media kept at room temperature into the mixture. <br/>
| |
− | <li>Incubate the mixture at 37 °C for 60 minutes </li>
| |
− | <li>Plate 50 ul of cells to the plate with appropriate antibiotic and incubate overnight at 37 °C</li>
| |
− | </div>
| |
| | | |
| <div style="height:80px"></div> | | <div style="height:80px"></div> |
| </div> | | </div> |
| </html> | | </html> |