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用于检测和定量印度香米(水稻)掺假的高通量多重微卫星标记分析

High-throughput multiplex microsatellite marker assay for detection and quantification of adulteration in Basmati rice (Oryza sativa).

作者信息

Archak Sunil, Lakshminarayanareddy V, Nagaraju Javaregowda

机构信息

Laboratory of Molecular Genetics, Centre for DNA Fingerprinting and Diagnostics, Nacharam, Hyderabad, India.

出版信息

Electrophoresis. 2007 Jul;28(14):2396-405. doi: 10.1002/elps.200600646.

Abstract

Basmati rice is a very special type of aromatic rice known world-wide for its extra long grains and pleasant and distinct aroma. Traditional Basmati rice cultivars, confined to Indo-Gangetic regions of the Indian subcontinent, are often reported to be adulterated with crossbred Basmati varieties and long-grain non-Basmati varieties in the export market. At present, there is no commercial scale technology to reliably detect adulteration. We report here a CE-based multiplex microsatellite marker assay for detection as well as quantification of adulteration in Basmati rice samples. The single-tube assay multiplexes eight microsatellite loci to generate variety-specific allele profiles that can detect adulteration from 1% upwards. The protocol also incorporates a quantitative-competitive PCR-based analysis for quantification of adulteration. Accuracy of quantification has been shown to be +/-1.5%. The experiments used to develop and validate the methodology are described.

摘要

巴斯马蒂大米是一种非常特殊的香米,以其超长的谷粒以及宜人且独特的香气而闻名于世。传统的巴斯马蒂水稻品种局限于印度次大陆的印度河—恒河地区,据报道,在出口市场上,它们常常被杂交巴斯马蒂品种和长粒非巴斯马蒂品种掺假。目前,尚无可靠检测掺假情况的商业规模技术。我们在此报告一种基于毛细管电泳的多重微卫星标记分析法,用于检测和定量巴斯马蒂大米样品中的掺假情况。这种单管分析法对八个微卫星位点进行多重分析,以生成品种特异性等位基因图谱,能够检测出1%及以上的掺假情况。该方案还纳入了基于定量竞争PCR的分析方法来定量掺假情况。定量的准确度已证明为±1.5%。文中描述了用于开发和验证该方法的实验。

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