[密花石斛的等位基因特异性诊断PCR鉴定]
[Allele-specific diagnostic PCR authentication of Dendrobium thyrsiflorum].
作者信息
Ying Yi, Xu Hong, Wang Zheng-Tao
机构信息
Key Laboratory of Standardization of Chinese Medicines of the Ministry of Education, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
出版信息
Yao Xue Xue Bao. 2007 Jan;42(1):98-103.
The aim of this study was to establish a diagnostic PCR method for authentication of D. thyrsiflorum Rchb. f. Based on rDNA ITS sequences of the 164 samples from 109 Dendrobium species sequenced and quoted from GenBank, the allele-specific diagnostic primers QH-JB1 and QH-JB2 for authentication were designed. Using the primers, diagnostic PCR was performed with the total DNA of D. thyrsiflorum Rchb. f. and other D. species as templates to establish the positive PCR condition for D. thyrsiflorum Rchb. f. A DNA fragment about 300 bp was amplified from D. thyrsiflorum Rchb. f. with anneal temperature at 63.5 degrees C and anneal time at 1 min, whereas no other DNA fragment was amplified from the rest D. species. The allele-specific diagnostic PCR was proved to be a simple and quick identification technique, suitable for the authentication of both fresh and dried materials of D. thyrsiflorum Rchb. f.
本研究的目的是建立一种用于鉴别鼓槌石斛（D. thyrsiflorum Rchb. f.）的诊断性聚合酶链反应（PCR）方法。基于从109种石斛属植物中测序并从GenBank引用的164个样本的核糖体DNA内转录间隔区（rDNA ITS）序列，设计了用于鉴别的等位基因特异性诊断引物QH-JB1和QH-JB2。使用这些引物，以鼓槌石斛的总DNA和其他石斛属植物的总DNA为模板进行诊断性PCR，以建立鼓槌石斛的阳性PCR条件。在退火温度为63.5℃、退火时间为1分钟的条件下，从鼓槌石斛中扩增出一条约300 bp的DNA片段，而其余石斛属植物未扩增出其他DNA片段。等位基因特异性诊断PCR被证明是一种简单快速的鉴定技术，适用于鼓槌石斛新鲜材料和干燥材料的鉴别。
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