ligation of PCR products
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.
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