Sequence-specific detection of PCR-amplified DNA by restriction enzyme release of hybrids

被引：2

作者：

Estrada, G ^{[1
]}

Colin, L ^{[1
]}

Gaytan, P ^{[1
]}

Alagon, A ^{[1
]}

Lizardi, PM ^{[1
]}

机构：

[1] UNIV NACL AUTONOMA MEXICO,INST BIOTECNOL,DEPT MOL RECOGNIT & STRUCT BIOL,CUERNAVACA 62271,MORELOS,MEXICO

来源：

MOLECULAR AND CELLULAR PROBES | 1996年 / 10卷 / 03期

关键词：

PCR; microtitre; nucleic acid detection; fluorescence; Entamoeba histolytica; repetitive sequences;

D O I：

10.1006/mcpr.1996.0025

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We have developed a method to detect PCR amplicons that contain a restriction enzyme site. The method is called Polymerase Chain Reaction-Hybridization and Enzymatic Release Assay (PCR-HERA). The PCR product is hybridized to a synthetic oligonucleotide that is immobilized on a microtitre plate. Specific cleavage of the resulting hybrid by a restriction endonuclease liberates a fluorescein labeled arm of the immobilized DNA. We demonstrate the use of PCR-HERA to detect species-specific sequences of extra-chromosomal, ribosomal DNA, of Entamoeba histolytica. This highly specific assay does not require any washing steps and can be easily automated. (C) 1996 Academic Press Limited

引用

页码：179 / 185

页数：7

共 12 条

[1] SENSITIVE AND QUANTITATIVE DETECTION OF PCR-AMPLIFIED HIV-1 DNA PRODUCTS BY AN ENZYME-LINKED IMMUNOASSAY FOLLOWING SOLUTION HYBRIDIZATION WITH 2 DIFFERENTLY LABELED OLIGONUCLEOTIDE PROBES [J].