Mutou Chiaki, Tanaka Katsunori, Ishikawa Ryuji
National Institute of Aerobiological Sciences, Tsukuba, Ibaraki, Japan.
Methods Mol Biol. 2014;1099:7-15. doi: 10.1007/978-1-62703-715-0_2.
Deoxyribonucleic acid (DNA) extracted from endosperm can be effectively used for rapid genotyping using seed tissue, to evaluate seed quality from packaged grains and to determine the purity of milled grains. Methods outlined here are optimal procedures to isolate DNA from endosperm tissue of modern rice grains and of aged rice remains preserved between 50 and 100 years. The extracted DNA can be used to amplify regions of chloroplast genomic DNA (ctDNA), mitochondrial genomic DNA (mtDNA), and nuclear genomic DNA using standard PCR protocols. In addition, we describe an optimal procedure to process archaeological grain specimens, aged for a couple of thousand years, to isolate DNA from these ancient samples, referred to here as ancient DNA (aDNA). The aDNA can be successfully amplified by PCR using appropriate primer pairs designed specifically for aDNA amplification.
从胚乳中提取的脱氧核糖核酸（DNA）可有效地用于利用种子组织进行快速基因分型，以评估包装谷物的种子质量，并确定碾磨谷物的纯度。这里概述的方法是从现代水稻籽粒和保存50至100年的陈年水稻遗存的胚乳组织中分离DNA的最佳程序。提取的DNA可用于使用标准PCR协议扩增叶绿体基因组DNA（ctDNA）、线粒体基因组DNA（mtDNA）和核基因组DNA区域。此外，我们描述了一种处理有数千年历史的考古谷物标本的最佳程序，以便从这些古代样本（这里称为古DNA（aDNA））中分离DNA。使用专门为aDNA扩增设计的合适引物对，通过PCR可以成功扩增aDNA。
