Difference between revisions of "Team:UMaryland/Notebook2"
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+ | <div id='contentbox'> | ||
+ | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
+ | <b>Ligation</b></a> | ||
+ | <p style="font-size:24px;text-align:left;text-decoration: underline;"> | ||
+ | Materials: | ||
+ | <ul class="a"> | ||
+ | <li>- 2 µL PSBIA3 digest</li> | ||
+ | <li>- 2 µL upstream digest (pBAD/ sRNBC)</li> | ||
+ | <li>- 2 µLdownstream digest (miraculin / const_GFP)</li> | ||
+ | <li>- 1 µL T4 DNA ligase</li> | ||
+ | <li>- 2 µL T4 DNA ligase 10x rxn buffer</li> | ||
+ | </ul> | ||
+ | <p style="font-size:24px;text-align:left;text-decoration: underline;"> | ||
+ | Procedure: | ||
+ | <ul class="a"> | ||
+ | <li>- Combine reagents in a clean PCR tube</li> | ||
+ | <li>- Let stand 10 mins at room temperature/ no heat kill</li> | ||
+ | |||
+ | </ul> | ||
+ | <br> | ||
+ | <br> | ||
+ | </div> | ||
+ | <div id='contentbox'> | ||
+ | <p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;"> | ||
+ | <b>Transformation</b></a> | ||
+ | <p style="font-size:24px;text-align:left;text-decoration: underline;"> | ||
+ | Materials: | ||
+ | <ul class="a"> | ||
+ | <li>- 50 µL cells (DH5alpha)</li> | ||
+ | <li>- 2 µL DNA </li> | ||
+ | </ul> | ||
+ | <p style="font-size:24px;text-align:left;text-decoration: underline;"> | ||
+ | Procedure: | ||
+ | <ul class="a"> | ||
+ | <li>- Add DNA to cells</li> | ||
+ | <li>- Incubate on ice for 30 minutes</li> | ||
+ | <li>- Heat shock at 42° fro 30 seconds</li> | ||
+ | <li>- Add 1 mL SOC media to the cells</li> | ||
+ | <li>- Incubate for 60 minutes at 37°</li> | ||
+ | <li>- Plate 200 µL </li> | ||
+ | <li>- Incubate at 37°</li> | ||
+ | |||
+ | </ul> | ||
+ | <br> | ||
+ | <br> | ||
+ | </div> | ||
</div> | </div> | ||
</html> | </html> |
Revision as of 10:13, 18 September 2015