Team:UT-Dallas/Notebook/8-22

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<LI> B3 reporter vectors on Chlora: Inoculate
<LI> B3 reporter vectors on Chlora: Inoculate
<LI> B4 reporters-chlora: Glycerol stock + Miniprep + digest + gel extract + purify + ligate + transform onto carb back bone.
<LI> B4 reporters-chlora: Glycerol stock + Miniprep + digest + gel extract + purify + ligate + transform onto carb back bone.
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<LI> Inoculate tcpA(3)-chlora reporter.
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<LI> Inoculate acfA(3)-chlora reporter.
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Revision as of 06:21, 24 August 2014

Friday, August 22, 2014

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Today is also long day. (We almost repeated everything we did yesterday: inoculation, miniprep, ligation, and transformation. Also, sequencing results are here.

Second batch (B2):

  • toxT: got very high noise in the background. We are possibly re-sequencing them.

  • tcpA: (gRNA+Promoter) first clone was wrong after sequencing. We have two clones so we will re-test-digesting-2%-gel them and send them out for sequencing.

Third batch (B3): Yesterday, we transformed reporter-Carb; today, we inoculate them. The colonies are not very big, we have about 3 colonies on the negative; however, the other plates are very good: they has at least twice-fold the colonies on the negative.

Forth batch (B4): We inoculated yesterday. We are minipreping, digesting, ligating onto Carb back bone, and transforming them

tcpQ is being re-digested and re-tested on 2% gel to make sure of our clone. We tested it on 1% gel but it created a smear and was dubious.


Ran out of PstI-HF!!! Need to order!!!


Tomorrow: For B4 reporters and B3 gRNA, inoculation. B3 reporters: miniprep. Sunday, we will miniprep. Next week, on Monday, we should have everything ready for test digest and then finally send the last batch of sequencing!

Today's tasks:

  • B3 reporter vectors on Chlora: Inoculate
  • B4 reporters-chlora: Glycerol stock + Miniprep + digest + gel extract + purify + ligate + transform onto carb back bone.
  • Inoculate acfA(3)-chlora reporter.


 
Terrible, terrible gel! We punctured the gel! The lane bled over each other. We had to make another gel to purify.
(In order, i5.47.1-2-3-4-X-5-6-7-8-9-10-11-12)

Updating the wiki notebook from the slowest computer in the building because currently we can't log into any other computer. Rishika is gel-ing the ctx-s.