Novel chemical method for the preparation of nucleic acids for nonisotopic hybridization

R. P. Viscidi, C. J. Connelly, R. H. Yolken

Research output: Contribution to journalArticlepeer-review

Abstract

A novel chemical method was used to prepare biotin-labeled nucleic acids for nonisotopic hybridization. The method involes the transamination of unpaired cytosine residues in polynucleotides with sodium bisulfite and ethylenediamine. Primary amino groups on the cytosine derivatives are then reacted with biotinyl-e-aminocaproic acid N-hydroxysuccinimide ester. Biotinylated probes hybridized with 1 to 2 pg of nitrocellulose filter-bound DNA and were visualized with a colorimetric detection technique. This method is simpler and less expensive than other methods for the preparation of nonisotopic probes. In addition, it is more versatile since the chemically modified bases can potentially react with other 'indicator' molecules or proteins such as an enzyme. The specificity for unpaired cytosine residues is another advantage which could allow for the selective labeling of a specific region of a double-strand nucleic acid. This improved labeling method should lead to the wider application of hybridization techniques in diagnostic microbiology and basic research in infectious diseases.

Original languageEnglish (US)
Pages (from-to)311-317
Number of pages7
JournalJournal of clinical microbiology
Volume23
Issue number2
DOIs
StatePublished - 1986

ASJC Scopus subject areas

  • Microbiology (medical)

Fingerprint Dive into the research topics of 'Novel chemical method for the preparation of nucleic acids for nonisotopic hybridization'. Together they form a unique fingerprint.

Cite this