- International Journal of Systematic and Evolutionary Microbiology
- Volume 39, Issue 3
f 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
- Authors: TAKAYUKI EZAKI*, YASUHIRO HASHIMOTO, EIKO YABUUCHI
- *Corresponding author.
- First Published Online: 01 July 1989, International Journal of Systematic and Evolutionary Microbiology 39: 224-229, doi: 10.1099/00207713-39-3-224
- Subject: Original Papers Relating To Systematic Bacteriology
- Published Online:
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, Page 1 of 1< Previous page | Next page > /docserver/preview/fulltext/ijsem/39/3/ijs-39-3-224-1.gif
Fluorometric hybridization in microdilution wells was developed to determine genetic relatedness among microorganisms. Total chromosomal deoxyribonucleic acid (DNA) for hybridization reactions was labeled with photoreactive biotin (photobiotin). The biotinylated DNA was hybridized with single-stranded unlabeled DNAs which had been immobilized on the surfaces of microdilution wells. After hybridization, biotinylated DNA was quantitatively detected with beta-D-galactosidase and a fluorogenic substrate, 4-methylumbelliferyl-beta-D-galactopyranoside. Homology values obtained with this fluorometric direct binding method were compared with values obtained with two membrane filter methods, one in which photobiotin labeling was used and one in which radioisotope labeling was used. The results showed that the fluorometric direct binding method in which microdilution wells are used could be an alternative to radioisotope and membrane filter hybridization methods.
Copyright © 1989 International Union of Microbiological Societies | Published by the Microbiology Society
Article metrics loading...
Full text loading...
Author and Article Information
/content/journal/ijsem/10.1099/00207713-39-3-224dcterms_title,dcterms_subject,pub_serialTitlepub_serialIdent:journal/ijsem AND -contentType:BlogPost104
/content/journal/ijsem/10.1099/00207713-39-3-224dcterms_title,dcterms_subject-pub_serialIdent:journal/ijsem AND -contentType:BlogPost104
Figure data loading....