Team:Brown/Notebook/September24

From 2010.igem.org

Friday, September 24, 2010

Checking on Overnights

Came in to check on Ethan's overnights- looks like nothing grew - 8:15 AM. Perhaps due to selecting satellite colonies mistakenly? Nothing to miniprep.

Other Notes

Took plates from incubator at 9:15 AM

  • Tat-L in pSB1A3 plate: 5-8 colonies, will overnight later today.

(8:40 PM: overnight started)

  • Tat-L in pGEM: TONS of colonies (not countable); only a small fraction are blue! Will overnight later today.

(8:40 PM: two overnights started)

Retrieved PCR samples from thermocycler, will run on gel later

  • Look for ~100 bp segment amplified by Tat-fwd and rev primers. If present, confirm insertion of Tat-L into pBlueScript.

Miniprepped overnights from 9/23

  • labeled "mini #1 JH 9/24"

Overnight Cultures

15 overnights of big colonies of AraC, CI, Mnt as before.

  • Colony PCR on corresponding colonies

50 μl total:

  • 25 μl mix
  • 1.5 each primer (2x1.5) -- later realized this was 10x too much primer
  • 22 μl dH2O

PCR started at 1:38 PM.

5:35 PM

Put 15 culture tubes in the incubator.

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