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

Hi,
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.

check this post
http://www.protocol-online.org/forums/inde...&st=0&#entry827

http://www.protocol-online.org/forums/inde...&st=0&#entry506