i have run into a few problems in the experiment for plasmid extraction. i did all the steps for boiling lysis followed by phenol-chloroform protocol. for two tubes,i also gave an additional RNase digestion (incubation time 1hr @ 37C). during electrophoresis only the samples with phenol-chloroform treatment had the proper bands while the ones with phenol chloroform+RNase didn’t show any bands at all. can anybody help me interpret these observations?
Controls are your friend:
Did you have DNA in the tube in the first place (saving a few µl before digestion would have told you that)?
Is your RNAse not degrading DNA (test with a solution that you know contains DNA) because of a contamination?
ya i ran a few controls and observed that everything, even the standards when treated with RNase were showing up as a blank. i think there is some contamination in the RNase but can you explain why the entire lane is appearing as a blank?