I am doing DNA FISH on mice stem cells to detect an endogenous gene location. The expected result should have two signal spots, representing the two copies of the allele. However, what I see is multiple signal spots in my cells. Has anyone seen this issue before? If so, how should I change my experiment condition?
I am doing denature at 80C 20min-30min.
I also tried to increase the cot1 DNA(double the amount that I use), however, it didn't have too much improvement.
Thanks in advance!
Frederic Lepretre
Hi Jing
first maybe you could reconsider your probe to be insufficiently specified to your target, or get less cells concentrated samples to avoid multi layers on your preparation (you can imagine cytogenetics as one 3D technique).
all the best
fred
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