Northern Hybridization - qestion about RNA degradation - need some clues (Oct/06/2005 )
just wanted to clear one point.
During Northern Hybridization, when we transfer RNA from gel to membrane we should take precautions as it may degrade, but when transferred, is the RNA on membrane equally susceptible to degradation?
After transfer of RNA to membrane can we proceed further (prehybridization,hybridization, stringent washing, detection)without taking care of RNases or we must proceed under RNase free conditions till the end?
plzz explain in detail.
Once the RNA has been fixed to the membrane (by baking or stratalinking) you no longer have to worry about degradation or use RNAse free buffers etc... The RNA is still susceptible to degradation, but it is already attached to the membrane at the correct molecular weight etc. and nicking will not affect the results.
[quote name='beccaf22' date='Oct 6 2005, 12:52 PM' post='26996']