Double RE digest: using two different companies - (Apr/06/2007 )
How horrible is it to do a double enzyme digest with two enzymes from different companies?? I'm doing a NotI and EcoRI digest. Both companies suggest using EcoRI buffer in a double digest situation so I've been using the EcoRI's company's EcoRI buffer. I feel like this is a simple question but my digests aren't working and I'm trying to discount every possibility.
Thanks!!
compare the composition of the ecor1 buffers from the two companies then compare the composition of the not1 buffer from the company that supplied it to you.
I have done double digest with enzymes from 2 companies and used buffer from one of them only. Since one of them was from NEB, i used neb buffer. I didnt have any problem.
Verify separately if the enzymes digest properly in the respective buffers.
i regularly perform double digestion and enzymes from various company doesnt bother me. i use the common buffer for the double digestion from one company though i use the enzymes from two differnent company.
u can check the following link
http://www.neb.com/nebecomm/DoubleDigestCalculator.asp