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different restriction enzymes different companies, can it work?? - (Sep/19/2013 )

Hi all,

I have cloned xho1 and sal1 restriction sites into a gene and transformed it, i am now trying to check if the insert is there by digesting the extracted plamid after transformation, digesting it with the same 2 enzymes as said before and running on gel. i am used to use the enzymes from roche and there they have same buffer for both so it was easier. Now i have xho1 from roche and sal1 from neb, different companies, different buffers. i know it has to be sequential digestion, but worried about the certainity it would work. anyone done this before, did it work, any advice or suggestions?? thanks a lot in advance.


It will certainly work with each in their supplied buffer. It would likely work with both at the same time in a carefully selected buffer.