Ligating large fragment into vector
jimmy barbee
jlb at lilly.com
Wed Jun 26 12:21:28 EST 1996
I have been having trouble ligating a 4 Kb PCR fragment into a 4 Kb
vector. Whether into a T/A vector from the PCR reaction or after cutting
to leave sticky ends into another vector, I haven't been able to get a
single insertion. Can anyone suggest modified ligation reaction
conditions to help? I suspect a site on one of the ends isn't cutting
well or at all, but I don't know why the T/A approach didn't work.
Thanks,
Jimmy
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