Help about methylation and SexAI

Rich Dudley rdudley+ at pitt.edu
Mon Mar 15 08:52:19 EST 1999


Gianluca Molla wrote:

> Hi everyone,
> I need to use the SexAI restriction enzyme. On the NEB catalog
> is stated that this enzyme DO NOT cuts if its reconition
> site is methylated by dcm methylase. Now I think I have
> to purify my DNA from a E.coli strain that is dcm-, but
> almost every cloning strain in cdm+ (except JM110).
> Can I use this strain or its genotype is not good for cloning
> and plasmid amplification? And what about other methylases
> like EcoRII? Are they a problem?
> Maybe that my questions are quite stupid, but this is the first
> time I have a problem with methylation. SexAI is not very cheap,
> so I want to be sure that I'll be able to cut my DNA with it.
>
> Thank you for your attention.
>
> Gianluca

JM110 and SCS110 will work all right, but I have had much better sucecss
for all around cloning with NEB's GM2163 cells.  They'll give them to
you free upon request next time you order an enzyme from them.  The
catalog number is in th eback of their catalog.

Good luck!

rich

--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
http://www.cbp.pitt.edu
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