Vemireddy Lakshminarayana R, Archak Sunil, Nagaraju Javaregowda
Laboratory of Molecular Genetics, Centre for DNA Fingerprinting and Diagnostics, ECIL Road, Nacharam, Hyderabad 500 076, Andhra Pradesh, India.
J Agric Food Chem. 2007 Oct 3;55(20):8112-7. doi: 10.1021/jf0714517. Epub 2007 Sep 15.
Microsatellite markers are employed for genotyping of Basmati varieties and assaying purity of market samples. However, employment of diverse electrophoresis techniques across laboratories has resulted in inconsistent allele sizes, creating doubts about the suitability of the assay. This study evaluated agarose gel electrophoresis, slab gel electrophoresis, and capillary electrophoresis techniques for their efficiency in the detection and quantification of adulteration in Basmati samples. Comparative analysis across 8 microsatellite loci in 12 rice varieties demonstrated that the capillary electrophoresis method showed less error (+/-0.73 bp) in the estimation of allele sizes compared to slab gel (+/-1.59 bp) and agarose gel (+/-8.03 bp) electrophoretic methods. Capillary electrophoresis showed greater reproducibility (<0.5 bp deviation) compared to slab gel (1 bp) and agarose (>3 bp) based methods. Capillary electrophoresis was significantly superior in quantification of the adulterant, with a mean error of +/-3.91% in comparison to slab gel (+/-6.09%). Lack of accuracy and consistency of the slab gel and agarose electrophoretic methods warrants the employment of capillary electrophoresis for Basmati rice purity assays.
微卫星标记用于巴斯马蒂品种的基因分型和市场样品纯度检测。然而，不同实验室采用不同的电泳技术导致等位基因大小不一致，引发了对该检测方法适用性的质疑。本研究评估了琼脂糖凝胶电泳、平板凝胶电泳和毛细管电泳技术在检测和定量巴斯马蒂样品掺假方面的效率。对12个水稻品种的8个微卫星位点进行的比较分析表明，与平板凝胶电泳（±1.59 bp）和琼脂糖凝胶电泳（±8.03 bp）方法相比，毛细管电泳法在等位基因大小估计上的误差较小（±0.73 bp）。与基于平板凝胶（1 bp）和琼脂糖（>3 bp）的方法相比，毛细管电泳显示出更高的重现性（偏差<0.5 bp）。在掺假物定量方面，毛细管电泳明显更优，平均误差为±3.91%，而平板凝胶电泳为±6.09%。平板凝胶电泳和琼脂糖电泳方法缺乏准确性和一致性，因此在巴斯马蒂大米纯度检测中应采用毛细管电泳。
