Cloning PCR products
aiyar at ebv.oncology.wisc.edu
Tue Aug 20 09:32:51 EST 1996
On Mon, 19 Aug 1996 10:40:13 GMT, Dr Andrew J. Doherty <doherty at bsa.bristol.ac.uk> wrote:
>THis is a very basic, even silly question. But even so, I'm trying to
>clone a series of PCR products. Do I need to kinase those products prior
>to ligation? The vector is de-phophorylated. It seems logical that I
>should, but then shouldn't the primers have 5'phosphates any way?
You should either kinase the PCR products, or better yet, the oligo-
nucleotides prior to PCR. Unless you specifically requested the
addition of 5'-phosphates, oligonucleotides are synthesized with
a 5'-hydroxyl group.
Ashok Aiyar, Ph.D.
Department of Oncology email: aiyar at ebv.oncology.wisc.edu
University of Wisconsin-Madison tel: (608) 262-6697
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