Development of a SYBR safe technique for the sensitive detection of DNA in cesium chloride density gradients for stable isotope probing assays.
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.
Martineau
Christine
C
Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Ave., Montreal, QC, Canada H4P 2R2.
Whyte
Lyle G
LG
Greer
Charles W
CW
D002247:Carbon Isotopes / Q000378:metabolism; D002499:Centrifugation, Density Gradient / Q000379:methods*; D002586:Cesium; D002712:Chlorides; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods*; D012680:Sensitivity and Specificity
D002247:Carbon Isotopes; D002712:Chlorides; D005456:Fluorescent Dyes; D002586:Cesium; D004247:DNA; C028019:cesium chloride