ligation of PCR products

Chi-kuang Wen ckwen at expert.cc.purdue.edu
Thu Oct 17 09:44:36 EST 1996


Gary Kucera (kucera~gt at glaxo.com) wrote:
: Zhiyong Zhao wrote:
: > 
: > I wonder if it is possible to add phosphate group to the 5'-end in the primer s
: > ynthesis reaction.


: Someone correct me if I am wrong because I am going under 
: the assumption that if an oligo has been de-protected 
: following synthesis, it SHOULD contain a 5' phosphate group.  
: Oligos are synthesised in a 3'->5' direction, therefore, a 
: phosphate group Must be present on the last base added (i.e. 
: de-protected) before the next base is added.
: -- 
: ________________________________________________________
: Gary T. Kucera, Scientist
: Cellular Sciences/Transgenics Group
: GlaxoWellcome Research, Inc.
: Five Moore Drive
: Research Triangle Park, NC 27709

The primers may be phosphorylated by T4 polynucleotide kinase at
the presence of ATP. The synthesized primers DO NOT have 5- phosphate.

Chi-Kuang
 



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