用时间分辨荧光法对硝酸纤维素膜上的核酸进行定量分析。

Christopoulos T K, Diamandis E P, Wilson G

Department of Clinical Biochemistry, Toronto Western Hospital, Ontario, Canada.

Nucleic Acids Res. 1991 Nov 11;19(21):6015-9. doi: 10.1093/nar/19.21.6015.

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.

我们使用一种基于链霉亲和素的大分子复合物（SBMC）作为染色试剂，该复合物用4,7-双（氯磺苯基）-1,10-菲咯啉-2,9-二羧酸（BCPDA）的铕螯合物进行标记，用于检测硝酸纤维素滤膜上生物素化的DNA。所获得的荧光斑点或条带既可以在紫外光照射下观察，用即时相机拍照，也可以使用专门设计的作为高分辨率时间分辨荧光扫描仪的仪器进行定量。检测限约为10 pg的目标DNA。使用与Southern转移靶标杂交的生物素化DNA探针进行的各种实验表明，该新方法是一种用于在固体支持物上染色DNA的有用的通用非同位素方法。