Cloning PCR products

Ashok Aiyar 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
-- 
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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