Team:BYU Provo/Team Thermosensor/Week14


Team BYU Provo

BYU Provo


Week 14 (Jul. 17-23)


Sequencing results back

pIG 15-IG40 (396bp)--aligned with lacZ from pcDNA3_lv5hislacz_seq

  • score 250 bits
  • identities 164/188 (87%)
  • Strand plus/plus
  • E=4.8e^-70

pIG16-IG40 (640bp)--aligned with lacZ from pcDNA3_lv5hislacz_seq

  • Score 1029 bits
  • E=0

pIG17-IG12 (963bp)--aligned with sfGFP sequence from Dr. Bundy

  • No significant match

pIG 15 and pIG16 have inserted correctly! We can go ahead and move forward with those!

pIG17 & most likely pIG18 don’t have GFP



Boiled template for LacZ∆TS (setting up to check for thermosensor insertion)

selected 8 colonies, diluted in 50microliters ddH20, then streaked on 8-slice plate (41-1). Also streaked for GFP∆TS (41-2)



Standard Taq PCR set up as colony PCR for 8 colonies from plates 41-1 and 42-1.

  • Plate 41-1 colony PCR only because 41-2 colony PCR got “royally messed up” Not certain of the accuracy.


Remade boiled template so we can redo colony PCR if needed

  • T41-1 boiled template M
  • T41-2 boiled template M

Ran Gel on previously set up colony PCR--there are two bands, one looks like possible thermosensor, the other is unknown

Made XGAL/ARA/AMP plates:

  • 10g bacto-tryptone
  • 5g yeast
  • 5g NaCl
  • 5g Agar
  • .1g Arabinose
  • 1ml Amp
  • 1ml X-Gal

7 July 2011

Made LB/amp/ara/x-gal plates, put in fridge. Be CAEFUL: one bag has unlabeled plates. Label once dry.