Development of a SYBR safe technique for the sensitive detection of DNA in cesium chloride density gradients for stable isotope probing assays

Christine Martineau; Lyle G Whyte; Charles W Greer

doi:10.1016/j.mimet.2008.01.016

Development of a SYBR safe technique for the sensitive detection of DNA in cesium chloride density gradients for stable isotope probing assays

J Microbiol Methods. 2008 May;73(2):199-202. doi: 10.1016/j.mimet.2008.01.016. Epub 2008 Feb 14.

Authors

Christine Martineau¹, Lyle G Whyte, Charles W Greer

Affiliation

¹ Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Ave., Montreal, QC, Canada H4P 2R2.

PMID: 18329741
DOI: 10.1016/j.mimet.2008.01.016

Abstract

SYBR safe, a fluorescent nucleic acid stain, was evaluated as a replacement for ethidium bromide (EtBr) in cesium chloride (CsCl) density gradients for DNA stable isotope probing (DNA-SIP) assays. The separation of 12C- and 13C-labelled DNA using SYBR safe gave similar results to those obtained using EtBr with pure cultures and environmental samples exposed to a 13C-labelled substrate, while the detection limit of DNA was enhanced by the use of SYBR safe by at least 5 times. The results demonstrated that SYBR safe is a safe, sensitive and effective alternative to the use of ethidium bromide in CsCl density gradients for DNA-SIP assays.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Carbon Isotopes / metabolism
Centrifugation, Density Gradient / methods*
Cesium
Chlorides
DNA / analysis*
DNA / isolation & purification*
DNA / metabolism
Fluorescent Dyes / metabolism
Isotope Labeling / methods*
Sensitivity and Specificity

Substances

Carbon Isotopes
Chlorides
Fluorescent Dyes
Cesium
DNA
cesium chloride