A rapid and sensitive determination of bacterial rRNA by means of hybridization-competition

✍ Scribed by Ben A. Oostra; Bert Zantinge; Aly L. van Goor; Albert J.J. van Ooyen; Max Gruber

• Publisher: Elsevier Science
• Year: 1976
• Tongue: English
• Weight: 446 KB
• Volume: 74
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(76)90231-1

✦ Synopsis

A rapid and sensitive liquid hyb~dization-competition method for the assay of in vitro synthesized Escherichia coli rRNA was developed. Hybridization with homologous DNA and with heterologous DNA from Proteus vulgaris was compared. Besides unfractionated DNA we used DNA enriched for the sites complementary to ribosomal RNA by chromatography on methylated albumin Kieselguhr. Hyb~dization with heterologous enriched DNA is the method of choice due to its low background and high hybridization efficiency.

Methods

Bacterial strains. E. coli CP78 (leu-, his-, arg-, three-, B1-, RCSfr) and Proteus v~lgar~s were used in this study.

Mqterials . Unlabeled nucleotides were from Boehringer, Mannheim, Germany; [5,6-3H]UTP (40-50 Ci/mmol) was from The Radiochemical Centre, Amersham, U.K. Bovine pancreatic ribonuclease was from 4%