1st experiment in 2007!!
Back in the year..2005 I have already selected Rif resistant kw20 mutants. As I recall mutant "1" & "i" have only a one point mutation and mutant "R1" & "R2" have two points mutation (confirmed by the Davies' lab) but resist to a higher concentration of Rif than 1 & i.
So here's the plan discussed with Rosie:
Make some overnight culture of 1 or i and R1 or R2 and wildtype kw20 (control) -> take 100ul into 20ml of sBHI -> grow until O.D. = 1.0 -> take 1ml from the flask into a test tube with Map7 DNA -> roll for 15' -> DNase 5' -> series of dilution -> plate on Nov plates sBHI plates.
Result: Compare the TF between RifR mutants & wildtype kw20
When to do this? some time in Jan!
Let me know if I should make change to this plan above. Thanks!!!