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Team:Duke/Notebook/August
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| + | <div class="day"> | ||
| + | <a id="aug8"><h2>August 8</h2></a> | ||
| + | <div class="obj">Objective: Backbone switching</div> | ||
| + | <div class="ppl">Delta Ghoshal, Matthew Faw</div> | ||
| + | <div class="lab"> | ||
| + | <p>Redo minipreps from yesterday - culture tubes grew overnight from salvaged cells</p> | ||
| + | <ul> | ||
| + | <li>pSB1C3-dCas9-tracrRNA - 171.3 ng/ul</li> | ||
| + | <li>pSB3C5-R0040-a-tracr 1 - 257.5 ng/ul</li> | ||
| + | <li>pSB3C5-R0040-a-tracr 2 - 202.7 ng/ul</li> | ||
| + | <li>pSB3C5-R0040-a-tracr 3 - 200.5 ng/ul</li> | ||
| + | <li>pSB3C5-R0040-a-tracr 4 - 198.3 ng/ul</li> | ||
| + | </ul> | ||
| + | |||
| + | <p>Analytical digest of Tube 1 (pSB1C3-dCas9-tracrRNA) using XhoI and BamHI</p> | ||
| + | <ul> | ||
| + | <li>2 hour digest at 37 degrees C</li> | ||
| + | <li>3X Master Mix:</li> | ||
| + | <ul> | ||
| + | <li>3 ul Cutsmart</li> | ||
| + | <li>22.5 ul water</li> | ||
| + | <li>0.75 ul XhoI</li> | ||
| + | <li>0.75 ul BamHI</li> | ||
| + | </ul> | ||
| + | <li>9 ul Master Mix added to 1 ul of pdCas9 in one tube and 1 ul of pSB1C3-dCas9-tracrRNA in another tube</li> | ||
| + | <li>Gel showed BLANK</li> | ||
| + | </ul> | ||
| + | </div> | ||
| + | </div> | ||
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Revision as of 22:12, 8 August 2014
August 8
Objective: Backbone switching
Delta Ghoshal, Matthew Faw
Redo minipreps from yesterday - culture tubes grew overnight from salvaged cells
- pSB1C3-dCas9-tracrRNA - 171.3 ng/ul
- pSB3C5-R0040-a-tracr 1 - 257.5 ng/ul
- pSB3C5-R0040-a-tracr 2 - 202.7 ng/ul
- pSB3C5-R0040-a-tracr 3 - 200.5 ng/ul
- pSB3C5-R0040-a-tracr 4 - 198.3 ng/ul
Analytical digest of Tube 1 (pSB1C3-dCas9-tracrRNA) using XhoI and BamHI
- 2 hour digest at 37 degrees C
- 3X Master Mix:
- 3 ul Cutsmart
- 22.5 ul water
- 0.75 ul XhoI
- 0.75 ul BamHI
- 9 ul Master Mix added to 1 ul of pdCas9 in one tube and 1 ul of pSB1C3-dCas9-tracrRNA in another tube
- Gel showed BLANK
