Nucleic Acids Research, Vol 25, Issue 14 2792-2799, Copyright © 1997 by Oxford University Press
J Weiler, H Gausepohl, N Hauser, ON Jensen and JD Hoheisel
Arrays of up to some 1000 PNA oligomers of individual sequence were
synthesised on polymer membranes using a robotic device originally designed
for peptide synthesis. At approximately 96%, the stepwise synthesis
efficiency was comparable to standard PNA synthesis procedures. Optionally,
the individual, fully deprotected PNA oligomers could be removed from the
support for further use, because an enzymatically cleavable but otherwise
stable linker was used. Since PNA arrays could form powerful tools for
hybridisation based DNA screening assays due to some favourable features of
the PNA molecules, the hybridisation behaviour of DNA probes to PNA arrays
was investigated for a precise understanding of PNA-DNA interactions on
solid support. Hybridisation followed the Watson-Crick base pairing rules
with higher duplex stabilities than on corresponding DNA oligonucleotide
sensors. Both the affinity and specificity of DNA hybridisation to the PNA
oligomers depended on the hybridisation conditions more than expected.
Successful discrimination between hybridisation to full complementary PNA
sequences and truncated or mismatched versions was possible at salt
concentrations down to 10 mM Na+and below, although an increasing tendency
to unspecific DNA binding and few strong mismatch hybridisation events were
observed.
ARTICLES
Hybridisation based DNA screening on peptide nucleic acid (PNA) oligomer arrays
Molecular-Genetic Genome Analysis, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 506, D-69120 Heidelberg, Germany.
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