RNAse protection assay
Posted 21 February 2005 - 11:02 PM
I'm doing a RNAse protection assay but I never see protected bands on the gel only background, even in the negative control i still have background. The sequence was sequenced twice, and the are antisense to the probe I want to see, the Tm of the shortest protected band was calculated (74°C) and I incubated the hybridisation mixture o.n. @ 65°C. And the probes are not degraded.
Did anyone have the same problems and could solve them finally?
Posted 22 February 2005 - 08:01 PM
PS We hybridize at 56c overnight.....but then again we use a purchased DNA template set
Posted 22 February 2005 - 08:08 PM