基于 Amplifluor 的 SNP 基因分型。

Amplifluor-Based SNP Genotyping.

机构信息

ICAR-Indian Institute of Oilseeds Research, Hyderabad, India.

出版信息

Methods Mol Biol. 2023;2638:191-200. doi: 10.1007/978-1-0716-3024-2_13.

Abstract

Amplifluor, a genotyping system used to analyze single nucleotide polymorphisms (SNPs), is supplied by Merck-Millipore. Amplifluor is based on polymerase chain reaction (PCR) with two competing allele-specific primers and a SNP specific common reverse primer. Sequence information flanking SNP of interest and fluorescent plate reader for end-point measurement or qPCR machine for real time measurement are required for the execution of the Amplifluor assay. In this chapter, the principle and working protocol of the Amplifluor assay based on end-point fluorescence detection of SNP allele is presented with an example.

摘要

Amplifluor 是一种用于分析单核苷酸多态性 (SNP) 的基因分型系统，由默克密理博公司提供。Amplifluor 基于聚合酶链反应 (PCR)，使用两种竞争性等位基因特异性引物和一个 SNP 特异性通用反向引物。执行 Amplifluor 分析需要与感兴趣的 SNP 侧翼的序列信息和终点荧光检测用荧光板读数器或实时测量用 qPCR 机器。本章介绍了基于 SNP 等位基因终点荧光检测的 Amplifluor 分析的原理和工作方案，并提供了一个实例。