Protocol Online logo
Top : Forum Archives: : Molecular Biology

Restriction digestion - (Mar/12/2007 )

Hi,
What would be the odd results due to setting up an overnight restriction digestion with ECO-R1

-avestha-

I dont think so unless you have high amount of Eco-RI to start with. Overnight digestion using that enzyme shouldnt give you any star activity. Again it depends on various factors... glycerol, amount of DNA and amount of enzyme used. wink.gif

-timjim-

I disagree

overnight incubation is certainly a way to get star activity with EcoRI. please see this page on NEB's site. EcoRI is subject to star activity.

if you use very minimal enzyme, you might be able to get away with overnight; but I would not chance it with an enzyme that efficient.

-aimikins-