Nucleic Acids Research, Vol 25, Issue 13 2657-2660, Copyright © 1997 by Oxford University Press
I Afonina, M Zivarts, I Kutyavin, E Lukhtanov, H Gamper and RB Meyer
The tripeptide 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3)
binds to the minor groove of DNA with high affinity. When this minor groove
binder (MGB) is conjugated to the 5'-end of short oligodeoxynucleotides
(ODNs), the conjugates form unusually stable hybrids with complementary DNA
in which the tethered CDPI3group resides in the minor groove. We show that
these conjugates can be used as PCR primers. Due to their unusually high
binding affinity, conjugates as short as 8-10mers can be used to amplify
DNA with good specificity and efficiency. The reduced length primers
described here might be appropriate for the PCR amplification of viral
sequences which possess a high degree of variability (e.g., HPV, HIV) or
for recent techniques such as gene hunting and differential display which
amplify multiple sequences using short primer pairs.
ARTICLES
Efficient priming of PCR with short oligonucleotides conjugated to a minor groove binder
Epoch Pharmaceuticals Inc., 1725 220th Street SE, #104 Bothell, WA 98021, USA. iafonina@epochpharm.com
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