Post-ligation restriction

Jean-Paul CADORET, Gie-ra Montpellier PDG-DRV-RA-DRIM, +33-6714-4707 jpcadore at ifremer.fr
Fri Jun 28 06:54:13 EST 1996


Hi netters,

Could somebody give me an explaination to this:

I blunt ligated a 1300 bp PCR insert using PCR script from stratagene.
Everything went OK with white colonies and so on.
After a midi prep of 4 colonies and restriction analysis, several restriction
sites are definitively reluctant to cutting. I checked the enzyme temperature
as well as buffers. It seems that this appears more on one side of the MCS.
Is the vector corrupted?
by my ligations?
Had somebody that broblem once ?

Thanks
JP



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