lashed, on 15 November 2011 - 12:19 PM, said:
i'm trying to figure out the problem with sequencing my insert..i have cloned a 3kb fragment into a vector of around the same size...inorder to confirm i tried a pcr by amplifying 5 fragments from this 3kb fragment....n i got the products of expected sizes..but when i try to sequence the whole plasmid using vector specific primers that should sequence only my insert..instead i get the vector sequence only...what could this problem be due to??
Did you put it on gel?
(put the vector with the insert on gel).
It does seem weird what you tell, but many things can go wrong, contaminated PCR or maybe you used the wrong primers or...)
If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.