blunt ligation problems

medbh at biovax.leeds.ac.uk medbh at biovax.leeds.ac.uk
Fri Aug 14 11:00:43 EST 1998


I have been having difficulty ligating a 250bp and 1.4kb PCR product into a pCAT and pGEM vector using promega's T4 DNA ligase, both as blunt products.  I have tried various insert:vector ratio's and have not been getting any successful clones.  I have been getting very few white colonies (pGEM) compared to blue colonies and all of these picked do not contain plasmid!  Could anyone offer some helpful suggestions or methods that have worked.
Thanks 



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