Team:BYU Provo/Team Thermosensor/Week13

Week 13 (7/10 - 7-16)

7/12/11

Restriction digest of insert and vector

∆TS insert 39-1

• 14 microliters ddH2O
• 5 microliters NEB buffer 2
• .5 microliters 100x BSA
• 30 microliters ∆TS PCR (31-4)
• 1.5 microliters ECORI
• 1.5 microliters HINDIII

pPLATBADGFP 39-2

• same as above except digesting 30 microliters of pIG17

PPLADBADLACZ

• same as above except digesting 30 microliters of pIG16

Put all above reactions at 37o for 40 min

Ran on a low melt gel. Bands appeared appropriate size for respective DNA -Julie

7/13/11

Set pIG16, pIG17, and pIG15 up to be sequenced. We are especially curious to see what the 300bp gene was that was inserted (see July 7th)

Set up two ligations. One with our thermosensor and pPLATBADLACZ, and another with pPLATBADGFP (unsure if we acutally have GFP inserted, sequencing submitted as check)

7/15/11

Transformation. Did two transformations one for our recently ligated pPLATBADLACZ∆TS (40-1), and one for pPLATBADGFP∆TS (40-2) respectively.

• 100 microliters of transformation 40-1 plated on plate 40-1. 100 microliters of transformation 40-2 plated on plate 40-2.

-Devin & Dallin

7/16/11

Both plates 40-1 and 40-2 had growth. Plates were parafilmed and placed in fridge.

-Devin & Dallin