3 replies to this topic

[Muktikant]

I have prepared plasmid (PET3a) in large scale and found lot of RNA contamination. But i am scared to RNAse treatment for i may loose some amount of plasmid DNA. Can i go ahead with RE digestion and ligation without RNA treatment. Because anyway i will be  running my REdigest in electrophoresis and following extraction before ligation.

Thanking in advance for you suggestions.

[milanoj]

If your RNase is clean you won't need to worry about loosing any of your plasmid. Are REs inhibited by RNA? THat's a good question. To avoid having to answer it just perform an RNase A treatment. No worries..

[Muktikant]

Milanoj,
THANX.
Muktikant

[irlogan]

You will almost certainly be safe treating your DNA with
RNaseA. Add an excess of the enzyme, then treat @ 37C for 1 hour. Phenol chloroform extract, then ethanol precipitate.

Works every time for DNA from Midi-preps etc etc

irl