Team:BYU Provo/Team Thermosensor/Week14

Week 14 (Jul. 17-23)

7/19/11

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

-Mark

7/19/11

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)

-Julie

7/20/11

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.

7/21/11

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.