Open Notebook: Cell cycle, MSH2, mutagen treatment

Thursday, January 8, 2015

Cell cycle, MSH2, mutagen treatment

A Gammie:

"We have never observed a growth defect with the null or any of the mutants, but we think there might be some differences in the presence of DNA damaging agents (HU or cisplatin) – you could try that for your flow experiments. I wonder if your strains have picked up secondary mutations. We never propagate the strains for very long because they are mutators and pick up deleterious mutations during propagation."

Gammie used HU and MMS in her 2013 DNA repair paper

http://www.sciencedirect.com/science/article/pii/S1568786412002819

HU was 0.1M = 100mM and MMS was 0.04% in this paper. Treatments were 1.5 hours (about one generation time for the BY lab strain).

Hydroxyurea (HU) (no respiratory risk label on Sigma)

AKA, hydroxycarbamide
http://en.wikipedia.org/wiki/Hydroxycarbamide

http://www.sigmaaldrich.com/catalog/product/sigma/h8627?lang=en&region=US
http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Product_Information_Sheet/2/h8627pis.pdf
FW: 76.06,
So, 1 gram can make 2M solution of 1000x2 /76.06 mL = 13.1/2 = 6.5 mL (This is 1000/6.5 mg/mL = 154 mg/mL)

"Hydroxyurea is freely soluble in water, to at least 50 mg/ml (5%). However, because hydroxyurea decomposes in the presence of moisture, aqueous solutions are probably not stable. It is recommended to store hydroxy-urea at 2-8 °C. Hydroxyurea should be stored in a dry atmosphere in airtight containers." (Sigma)