Inversion of in situ synthesized oligonucleotides: improved reagents for hybridization and primer extension in DNA microarrays.

Abstract:

:Oligonucleotides synthesized in array format suffer from contamination by truncated species. We have developed a method to invert DNA molecules in situ after completed synthesis. Reactive functions at the 5'-ends of the oligonucleotides are permitted to react with functions on the support before the 3'-ends are released, in effect reversing the orientation of full-length oligonucleotides, while any 5'-truncated molecules are lost. This strategy serves both to purify in situ synthesized reagents and to reorient the oligonucleotides, causing them to expose free 3'-hydroxyls. In situ inverted oligonucleotides can be used in assays based on DNA polymerase-assisted extension of immobilized primers, and we demonstrate their utility in minisequencing and in pyrosequencing.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Kwiatkowski M,Fredriksson S,Isaksson A,Nilsson M,Landegren U

doi

10.1093/nar/27.24.4710

keywords:

subject

Has Abstract

pub_date

1999-12-15 00:00:00

pages

4710-4

issue

24

eissn

0305-1048

issn

1362-4962

pii

gkc680

journal_volume

27

pub_type

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