Panax ginseng-specific sequence characterized amplified region (SCAR) marker for testing medicinal products
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To screen genetic polymorphisms of Panax ginseng, as well as those of Panax quinquefolium and Panax notoginseng, analysis of random amplified polymorphic DNA (RAPD) was performed using 120 random primers. Of the successful amplicons obtained, the Panax ginseng-specific RAPD marker C-12 was cloned into a TA vector and sequenced (GenBank access number KU553472). Based on the sequence analysis results, a pair of primers specific to C-12 was designed. Finally, a SCAR marker-based identification system for Panax ginseng was developed after optimization of the reaction conditions. Using this method, two positive bands were stably observed at 300 bp and 130 bp in 33 batches of Panax ginseng samples tested, while negative results were obtained for another 101 batches of samples, including Panax quinquefolium, Panax notoginseng, adulterants, and other medicinal herbs. Thus, we successfully developed a PCR-based method for rapid and effective identification of Panax ginseng, which can be effectively used for the protection and utilization of germplasm resources and identification of the origins of Panax ginseng samples.
Key wordsPanax ginseng random amplified polymorphic DNA (RAPD) sequence characterized amplified regions (SCAR) molecular identification
为了研究人参、西洋参和三七的基因多态性,使用120 条随机引物进行了随机扩增多态性DNA 分析。将筛选获得的人参特异性RAPD 标记C-12 进行T-A 克隆、测序(GenBank 登录号KU553472)。 根据序列分析结果,设计了一对特异性引物,经优化反应条件,建立了人参的SCAR 标记鉴别体系。 在所有的33 批人参标本中,均能稳定地获得约300 bp 和130 bp 的2 条阳性扩增带;在混伪品以及西 洋参、三七和其他中药材共计101 批标本中均为阴性扩增。因此,建立了一种快速、有效的能准确、 特异性地鉴别人参的PCR 方法,可用于人参的种质资源保护、利用及其基原鉴定。
关键词人参 随机扩增多态性DNA(RAPD) 特征序列扩增标记(SCAR) 分子鉴定
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