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PMID: 1923800 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The use of oligodeoxynucleotide probes in chaotrope-based hybridization solutions.

Nucleic acids research ·Vol. 19 ·No. 19 ·1991-10-11 ·Pages 5143-51

Van Ness J, Chen L

Abstract

Hybridization solutions containing chaotropes may be used to modulate the thermal stability (Tm or Td) of oligodeoxynucleotide (ODN) duplexes or hybrids over a 90 degrees C range. Modulation of Td allows formulation of hybridization solutions that permit ambient temperature hybridization using most combinations of probe length, probe composition, target type, and facilitates development of convenient and rapid assay formats. The conditions required to achieve ODN duplex fidelity, and optimal yields of hybridized product, are described for trichloroacetate, thiocyanate, guanidinium salts and other chaotropic salts. The effects of different solid supports on Td are described. Also, a method is presented that uses chaotropic compounds to reduce background arising from signal ODN probes in a sandwich assay hybridization format.

MeSH Terms
Base Sequence Guanidines/metabolism Kinetics Molecular Sequence Data Nucleic Acid Hybridization Oligonucleotide Probes/chemistry Sodium Chloride/chemistry Solutions/chemistry Temperature Thiocyanates/metabolism Trichloroacetic Acid/metabolism
Chemicals
Guanidines Oligonucleotide Probes Solutions Thiocyanates Sodium Chloride Trichloroacetic Acid thiocyanate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Van Ness J
MicroProbe Corporation, Bothell, WA 98021.
Chen L
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/19.19.5143
Published
1991-10-11
Pages
5143-51
Language
English
Region
England
NLM ID
0411011
PMCID
PMC328868
Subset
IM
Grants
NIDCR NIH HHS · 1 R43 DEO 7819-01 · United States
NIDCR NIH HHS · 1 R44 DEO 7819-02 · United States
External Links
PubMed source DOI: 10.1093/nar/19.19.5143 PMC Full Text View journal details
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