de Vere Natasha, Rich Tim C G, Trinder Sarah A, Long Charlotte
National Botanic Garden of Wales, Llanarthne, Carmarthenshire, SA32 8HG, UK,
Methods Mol Biol. 2015;1245:101-18. doi: 10.1007/978-1-4939-1966-6_8.
DNA barcoding uses specific regions of DNA in order to identify species. Initiatives are taking place around the world to generate DNA barcodes for all groups of living organisms and to make these data publically available in order to help understand, conserve, and utilize the world's biodiversity. For land plants the core DNA barcode markers are two sections of coding regions within the chloroplast, part of the genes, rbcL and matK. In order to create high quality databases, each plant that is DNA barcoded needs to have a herbarium voucher that accompanies the rbcL and matK DNA sequences. The quality of the DNA sequences, the primers used, and trace files should also be accessible to users of the data. Multiple individuals should be DNA barcoded for each species in order to check for errors and allow for intraspecific variation. The world's herbaria provide a rich resource of already preserved and identified material and these can be used for DNA barcoding as well as by collecting fresh samples from the wild. These protocols describe the whole DNA barcoding process, from the collection of plant material from the wild or from the herbarium, how to extract and amplify the DNA, and how to check the quality of the data after sequencing.
DNA条形码技术利用特定的DNA区域来识别物种。世界各地都在开展相关工作,为所有生物类群生成DNA条形码,并将这些数据公开,以帮助人们了解、保护和利用全球生物多样性。对于陆地植物而言,核心DNA条形码标记是叶绿体中编码区域的两个部分,即rbcL和matK基因的一部分。为了创建高质量的数据库,每一个进行DNA条形码分析的植物都需要有一份标本凭证,与rbcL和matK的DNA序列一同保存。数据的使用者还应能够获取DNA序列的质量、所用引物以及跟踪文件等信息。每个物种都应选取多个个体进行DNA条形码分析,以检查错误并考虑种内变异。全球的植物标本馆提供了丰富的已保存和已鉴定的材料资源,这些材料既可以用于DNA条形码分析工作,也可以通过从野外采集新鲜样本的方式来使用。这些实验方案描述了整个DNA条形码分析过程,从在野外或植物标本馆采集植物材料,到如何提取和扩增DNA,以及测序后如何检查数据质量。