DNAse activity in room temperature - (May/18/2009 )
Hello all;
In my procedure, I'd like to use DNAse in one step. The caveat is I shouldn't heat the sample; it should be at least at room temperature (I checked - heating in 37C makes my complex fall apart). I am using Ambion's Turbo DNAse to augment activity in my 200 mM salt buffer. How much will the activity drop? Will the enzyme work in room temperature? The original procedure goes only, like, "we added DNAse". Duh...
I don't know how much the activity will drop, but you should be able to use it at room temperature. I've never used the Ambion DNAse, but I know that there are plenty of protocols out there that use DNAse @ RT in temperature-sensitive experiments. ... I'd just increase the incubation time.
Cheers,
-Carlton
Telomerase on May 18 2009, 11:34 PM said:
In my procedure, I'd like to use DNAse in one step. The caveat is I shouldn't heat the sample; it should be at least at room temperature (I checked - heating in 37C makes my complex fall apart). I am using Ambion's Turbo DNAse to augment activity in my 200 mM salt buffer. How much will the activity drop? Will the enzyme work in room temperature? The original procedure goes only, like, "we added DNAse". Duh...
When we do cell lysis for protein harvesting, we incubate at Rt for 10-15 minutes.
Thanks, I'll try it right away when the mice arrive 