Open Notebook: Bio125, strains from Gammie lab

Tuesday, January 13, 2015

Bio125, strains from Gammie lab

Received on Jan 13, 2015

Yeast strains

AGY 75, yeast strain with pSH44 reporter plasmid

AGY124, yeast strain with pRS413 and pSH44 (This is the plasmid control)

AGY125, yeast strain with the wild type pMSH2 and pSH44 (This is the wildtype MSH2 control)

Ecoli strain with plasmid

AG372, pmsh2-H658R

AG421, pmsh2-A618V

Yeast Media:

To grow AGY125 and AGY124

SC-His-Trp
-HIS to keep pMSH2
-TRP to keep pSH44 reporter plasmid

To grow AGY75, SC-Trp

I added 2.5ul 88mg/ml histidine stock to 5ml SC-His-Trp for a final concentration of 44ug/ml based on Gammie protocol according to Dr. Kioko

For plate growth, I dripped 1X histitine to the SC-HIS-Trp plate for AGY75

Ecoli Media:
LB + amp because pRS413 has a beta-lactamase which offer resistance to beta-lactam antibiotics such as ampicillin and penicillin.

I added 5ul of 50 mg/mL ampicilin stock to 5ml LB.

Amplicilin, histidine stocks, LB and SC-His-Trp media are provided by Dr. Kioko.

20150113, 4pm, yeast liquid sample were at 30C shaker, and Ecoli strains at 37C shaker.
The yeast plate of AGY124 and 125 were at 30C incubator.
Plate AGY75 were left outside for histidine to dry.

20150114. 10am. Liquid growth are obvious for both yeast and Ecoli strains. Kioko froze the Ecoli in 20% glycerol as stocks.

Observation:
AGY124 and AGY125 grew on SD-Trp-His-Ura plates. They form pink colonies when speaded dense population.

2015 Feb 12, I finally obtained SD-Trp-Ura plates, and spread AGY75 strains from the glass-tube cultures.
2015 Feb 16. AGY75 grows well on SD-Trp-Ura plate (Problem: some colonies became pink, but some remain white after a while. ).