the problem on plasmid ligation and transformation

Austin So (Hae Jin) haejin at netinfo.ubc.ca
Tue Aug 10 16:04:00 EST 1999


I think you should be concerned about the compatibility of the PCR reaction
mixture with the REs you are using. I would have probably band isolated
*first* and then digested, then putting everything together for the ligation
reaction. The other issue is whether the RE site is too close to the end of
the PCR product.

Adding an extra cloning step (ie. using a TA vector first) is probably the
less problematic though...

Cheers

Austin

"Dr. D. Zha" wrote:

>  The length
> of the plasmid is around 6 Kb and the insert is around 1 kb, and recover
> the DNA bands after digestion from agarose gel by kit, and using T4 DNA
> ligase which bought from Boheringer,

--
---
Austin P. So (Hae Jin)

I.I.S.G.P.
Biotechnology Laboratory
University of British Columbia

E-mail: haejin at netinfo.ubc.ca

http://www.interchange.ubc.ca/haejin/index.html (under construction)





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