论文部分内容阅读
为建立白及药材的分子标记鉴定方法,该文提取白及及其混伪品药材的基因组DNA,利用PCR技术扩增r DNA ITS2片段,片段经双向测序、排序、比对,构建邻接树,查找SNP位点。结果显示,ITS2序列不仅能够有效地区分白及与其混伪品,而且存在可用作白及、黄花白及与其混伪药材鉴别的SNP位点。针对标记白及药材的SNP位点设计引物,通过筛选引物和优化特异性扩增条件后,获取引物BJ59-412F,BJ59-412R,HHBJ-225R在同一扩增条件下,可有效扩增白及、黄花白及,其产物长度分别约为350,520 bp,而混伪品则无PCR条带产生。该文所述引物和建立的反应条件可成功将白及、黄花白及与其混伪品药材进行同步鉴定,操作快速、简捷,结果可靠,可作为白及药材的快速分子鉴定方法。
In order to establish the method of molecular marker identification of white and medicinal materials, the genomic DNA of white and its adulterated medicinal materials was extracted. The rDNA ITS2 fragment was amplified by PCR and sequenced and sequenced by two-way sequencing to construct the adjacent tree, Find SNPs. The results showed that ITS2 sequence not only can effectively distinguish between white and its adulterants, but also has SNP sites that can be used for identification of white and yellow flowers and their pseudo-medicinal herbs. Primers were designed according to SNP loci of marker white and medicinal materials, primers BJ59-412F, BJ59-412R and HHBJ-225R were obtained by screening primers and optimizing specific amplification conditions, under the same amplification conditions, they could effectively amplify white and , Yellow and white, the length of their products were about 350,520 bp, while there is no PCR bands produced in the adulterants. The primers and the reaction conditions established in the article can be used for the simultaneous identification of white and yellow flowers and their adulterated medicinal materials. The method has the advantages of rapid, simple and reliable operation, and can be used as a rapid molecular identification method for white and medicinal materials.