Team:Duke/Notebook/August

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Revision as of 22:12, 8 August 2014 by Delta.ghoshal (Talk | contribs)

May 2014
Month 2 of Project
sun mon tue wed thu fri sat
1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31

June 2014
Month 3 of Project
sun mon tue wed thu fri sat
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30

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

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