Nucleic Acids Research, Vol 24, Issue 16 3142-3148, Copyright © 1996 by Oxford University Press
E Timofeev, SV Kochetkova, AD Mirzabekov and VL Florentiev
Four types of polyacrylamide or polydimethyl-acrylamide gels for
regioselective (by immobilization at the 3' end) of short oligonucleotides
have been designed for use in manufacturing oligonucleotide microchips. Two
of these supports contain amino or aldehyde groups in the gel, allowing
coupling with oligonucleotides bearing aldehyde or amino groups,
respectively, in the presence of a reducing agent. The aldehyde gel support
showed a higher immobilization efficiency relative to the amino gel. Of all
reducing agents tested, the best results were obtained with a
pyridine-borane complex. The other supports are based on an acrylamide gel
activated with glutaraldehyde or a hydroxyalkyl-functionalized gel treated
with mesyl chloride. The use of dimethylacrylamide instead of acrylamide
allows subsequent gel modifications in organic solvents. All the
immobilization methods are easy and simple to perform, give high and
reproducible yields, allow long durations of storage of the activated
support, and provide high stability of attachment and low non-specific
binding. Although these gel supports have been developed for preparing
oligonucleotide microchips, they may be used for other purposes as well.
ARTICLES
Regioselective immobilization of short oligonucleotides to acrylic copolymer gels
Joint Human Genome Program: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
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