Team:BYU Provo/Team Thermosensor/Week17

Week 17 (Aug. 7 - 13)

8/8/11

Redid overnights to freeze down pBADLAT and pBADLATlacZ

Moved Dr. Grose’s 4 slice plate to the fridge from the 37o to the fridge

8/9/11

IGM 29- T26-1 (col. 2) (pIG 12)

IGM 30- T26-1 (col. 5) (pIG 13)

IGM 31- T36-2 (col. 7) (pIG 15)

IGM 32- T36-2 (col. 3) ( pIG 16)

Boiled Template (T47-1) placed in freezer

Streaked plates T47-1,2, & 3 from Dr. Grose’s 4 colony plate

T47-1 placed in 37oC at 3:25 pm

T47-2&3 placed in 30oC at 3:25pm (2 plates because T47-2 might have gotten messed up)

These plates are to see if there is any difference in expression of lacZ at 30oC vs. 37o

--Mark

8/10/11

pBAD PCR, according to standard phusion protocol. Used primers IG37 and IG38

Looked at 37oC plate compared to 30oC plate. Thermosensor A is white on 30oC plate, but a very dark blue on the 37oC plate. Thermosensors B,C, and D show no apparant difference

Not sure why these thermosensors are acting differently.

Restreaked thermosensor A, and picked several more colonies to compare. Chose colonies and labeled them E-N. Double plated each of these colony streaks one in each 30oC and 37oC, Colony A was repeated as a control to the additional new colonies.

-Julie

8/11/11

Colonies A, I, and J white at 30oC, and blue at 37oC

Set up overnights of these colonies in 5ml LB-Amp. Incubating at 37oC in shaker overnight

-Addision

8/12/11

Plasmid preps of A, I, and J got messed up. Re-did with 5ml LB-Amp and put in 37oC shaker.

-Addison

Set up transformation with pIG16 (plasmid with everything but thermosensor) and DH5alpa cells. Doing this to make sure there is no leaky lacZ expression. Std. transformation protocol. (T49-1)

--Julie

8/13/11

Placed T49-1 and overnights A, I, and J in fridge

--Addison