QUANTIFICATION OF NUCLEIC-ACIDS ON NITROCELLULOSE MEMBRANES WITH TIME-RESOLVED FLUOROMETRY

被引：21

作者：

CHRISTOPOULOS, TK

DIAMANDIS, EP

WILSON, G

机构：

[1] TORONTO WESTERN HOSP,DEPT CLIN BIOCHEM,399 BATHURST ST,TORONTO M5T 2S8,ONTARIO,CANADA

[2] UNIV TORONTO,DEPT CLIN BIOCHEM,TORONTO M5G 1L5,ONTARIO,CANADA

[3] CYBERFLYOR INC,TORONTO M5T 1X4,ONTARIO,CANADA

来源：

NUCLEIC ACIDS RESEARCH | 1991年 / 19卷 / 21期

基金：

英国医学研究理事会;

关键词：

D O I：

10.1093/nar/19.21.6015

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We use a streptavidin-based macromolecular complex (SBMC) labelled with the europium chelate of 4,7-bis (chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA) as a staining reagent for biotinylated DNA present on nitrocellulose filters. The fluorescent spots or bands obtained can either be observed under UV illumination, photographed by instant camera photography or quantified by using a specially designed instrument working as a high resolution time-resolved fluorometric scanner. The detection limit is approximately 10 pg of target DNA. Various experiments with use of biotinylated DNA probes hybridized to Southern transferred targets have shown that the new procedure is a useful versatile non-isotopic methodology for staining DNA on solid supports.

引用

页码：6015 / 6019

页数：5

共 21 条

[1] CHEMI-LUMINESCENCE LIGHTS UP [J].