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
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
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) 

See cell cycle arrest protocol form

Wikipedia entry shows that hydroxurea is a WHO essential medicine, and used in some leukemia treatment.

Hydroxyurea (Hydroxycarbamide) ab142613 - Abcam
Claim 100mM stock solution in water or DMSO?
Tripathi2011Genetics:  HU cause G2/M arrest in yeast cells.


Methyl methanesulfonate (MMS) (respiratory risk, require facial mask)

Molar mass110.13 g/mol
So, 1 gram make 1000x1 /110.13 mL = 9.08 mL

Cisplatin (very expensive at Sigma)

Need to know to their dosage to conclude which one is actually more expensive to do experiments. 


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