一种在微孔板中检测特异性扩增PCR片段的快速方法。

A rapid method for detecting specific amplified PCR fragments in microtiter plates.

作者信息

Ortiz A, Ritter E

机构信息

CIMA-Granja Modelo, Vitoria-Gasteiz, Spain.

出版信息

Nucleic Acids Res. 1996 Aug 15;24(16):3280-1. doi: 10.1093/nar/24.16.3280.

DOI:10.1093/nar/24.16.3280

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC146076/

Abstract

A simple method is presented to circumvent laborious and time consuming electrophoretic separations of specific PCR amplification products. Specific target DNA is amplified using nucleotides labelled with DIG-dUTP or biotin-dCTP. The labelled PCR products are separated from unincorporated nucleotides or oligonucleotides by using a positively charged DEAE cellulose matrix. Amplification products are visualized directly in the matrix using immunoenzymatic methods or streptavidin-conjugated enzymes. The detection process can be carried out within 2 h, allows the processing of large sample sizes and can potentially be automated.

摘要

本文提出了一种简单的方法，以规避对特定PCR扩增产物进行费力且耗时的电泳分离。使用用DIG-dUTP或生物素-dCTP标记的核苷酸扩增特定的目标DNA。通过使用带正电荷的DEAE纤维素基质，将标记的PCR产物与未掺入的核苷酸或寡核苷酸分离。使用免疫酶法或链霉亲和素偶联酶直接在基质中观察扩增产物。检测过程可在2小时内完成，允许处理大量样本，并且有可能实现自动化。

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本文引用的文献

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A method combining immunocapture and PCR amplification in a microtiter plate for the detection of plant viruses and subviral pathogens.

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Quantification of polymerase chain reaction products by affinity-based hybrid collection.基于亲和的杂交捕获法对聚合酶链反应产物进行定量分析。

Nucleic Acids Res. 1988 Dec 9;16(23):11327-38. doi: 10.1093/nar/16.23.11327.

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Microplate hybridization of amplified viral DNA segment.扩增病毒DNA片段的微孔板杂交

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Quantitative analysis of polymerase chain reaction (PCR) products using primers labeled with biotin and a fluorescent dye.使用生物素和荧光染料标记的引物对聚合酶链反应（PCR）产物进行定量分析。

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