A colorimetric method for DNA hybridization

Abstract

A general method has been developed which allows crosslinks to be produced between proteins and single-stranded DNA. Such single-stranded DNA protein complexes have been tested for blot hybridization using two colorimetrically detectable enzymes, namely peroxidase and alkaline phosphatase, as the protein moiety of the probe. After hybridization and incubation with a substrate solution sequences complementary to the probe can be visualized directly without the need of tedious cytochemical sandwich methods. This procedure will detect target sequences, a few kilobases long, in the 1- to 5-pg range.

New Approaches to the Detection of Microbial Plant Pathogens

“…Covalent linking of an enzyme to single-stranded nucleic acids (e.g. oligonucleotides) which then can be used directly as hybridization probes (Renz and Kurz, 1984;Li etal., 1987;McLaughlin eral., 1987;Seriwatana et al, 1987). Again, the final stage of the procedure is to develop a colorimetric or other appropriate reaction product.…”

“…Again, the final stage of the procedure is to develop a colorimetric or other appropriate reaction product. Approximately 1-5 pg of target sequence can be detected (Renz and Kurz, 1984). However, the enzyme must survive the hybridization conditions.…”

Fluorometric Deoxyribonucleic Acid-Deoxyribonucleic Acid Hybridization in Microdilution Wells as an Alternative to Membrane Filter Hybridization in which Radioisotopes Are Used To Determine Genetic Relatedness among Bacterial Strains

“…However, to carry out most of these hybridization experiments, DNA must be labeled with radioactive substances by nick translation (18) or random primed labeling (9). Recent developments have made it possible to label DNA with nonradioactive materials without using enzymes (10,11,17). Biotinylation of DNA with photoreactive biotin (photobiotin) (10) is one of these recent developments.…”

“…Biotinylated DNA has been used for Southern and dot blot hybridizations (10,14). Hybridized DNA fragments are usually visualized with alkaline phosphatase or peroxidase color detection methods (10,14,17). We used photobiotinylated DNA for quantitative detection of DNA-DNA hybridization to determine genetic relatedness among microorganism for taxonomic studies (4, 5,19).…”

A note on four nonradioactive labeling systems for dot hybridization detection of potato viruses

“…The other Systems of labeling gave a lower level of sensitivity. However, the enhanced peroxidase chemiluminescence System (Amersham), involving direct labeling of DNA probe (Renz and Kurz 1984), was found to be the most convenient approach for largescale détection of spécifie nucleic acid sé-quences. Probe labeling with peroxidase is very short (10 min) and détection of probes after washings can be obtained within 60 min.…”