proteinase treatment - (Jun/22/2013 )
Dear all,
I am trying to collect DNA from samples that require a pretty long proteinase K treatment.
I was wondering if I could do this "incubation" overnight (+ 14 hours of incubation). Or would this perhaps damage the DNA?
The incubation happends at 57°C , so I wonder if this is bad? (since I always hear you need to keep DNA at low temperatures to keep it stable..)
thanks in advance
DNA is a pretty stable molecule, the major damage would result from DNases, which will be largely inactive at 57 and the proteinase K and salt conditions in the digest should also help inactivate the DNases. It is common practice to do DNA extractions from mouse tail tips overnight under conditions pretty similar to those you describe.
bob1 on Sat Jun 22 21:55:33 2013 said:
DNA is a pretty stable molecule, the major damage would result from DNases, which will be largely inactive at 57 and the proteinase K and salt conditions in the digest should also help inactivate the DNases. It is common practice to do DNA extractions from mouse tail tips overnight under conditions pretty similar to those you describe.
Ok, good to know.
The reason I asked is because I did read that sometimes they advice not to let it stand for more than 8-10 hours.