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pcr ligation - problems with ligation (Jan/20/2004 )

I have difficulties with ligating digested pcr products to vectors. The size of the pcr products are in the range of 1.5 kb and the vector used is ycp111. I would appreciate any suggestions...ThankU


I guess you have incorporated restriction sites into your PCR product. If this is the case, I would say that some enzyme doesn't cut well even you have 4-5 extra bases outside the restriction site. Provide more bases 5' to the restriction site.

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