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problems with NdeI restriction enzyme? - (Jun/03/2006 )

I am trying to cut out a piece of DNA with Eco Ri and NdeI enzymes from a vector. double digestion in NdeI buffer or digesting with one and then gel purifying and digesting with the other. I am sure that EcoRi is cutting but i have problems with NdeI. i looked at the NEB site for the limitations but that didn't solve my problem. is there anyone else having problems with that enzyme or had problems before with it?


NEB suggests double digestion in EcoRI buffer. Did you try that and it didn`t work? huh.gif


Nooo sad.gif
i have done double digestions in Nde I buffer. I am using promega enzymes and Nde I is more sensitive and less effective than EcoRi so i thought it might not cut in a buffer other than its own. anyway, thank you very much, i should learn to utilize NEB sites and suggestions unsure.gif