基于DNA熔解曲线技术的金银花种质鉴定Germplasm identification of Lonicerae Japonicae Flos by DNA melt curve analysis
陈媞颖;蒋超;袁媛;陈康;周骏辉;赵玉洋;黄璐琦;
CHEN Ti-ying;JIANG Chao;YUAN Yuan;CHEN Kang;ZHOU Jun-hui;ZHAO Yu-yang;HUANG Lu-qi;State Key Laboratory of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;
摘要(Abstract):
将高分辨率熔解曲线(high resolution melting curve analysis,HRM)技术应用于金银花种质鉴定中,对已知7对SSR(简单重复序列,simple sequence repeats)引物进行筛选,通过熔解曲线峰型及Tm差异,确定适合鉴别引物并考察其DNA模板浓度、退火温度及循环数的适宜范围。结合SIMCA-P软件对荧光信号进行处理分析,结果表明利用其中4对引物的熔解曲线可以区分金银花的3个主栽种质,为进一步完善金银花种质鉴定方法提供了依据。
To establish a new high resolution melting analytical method for identification of Lonicera japonica germplasm,the screening of 7 pairs of SSR( simple sequence repeats) primers,determining the suitable diagnostic primers by the differences of peak pattern and Tmwas conducted. Then into the DNA template concentration,annealing temperature and the suitable range of cycle number were investigated. Combined with SIMCA-P software for data processing analysis,the results show that three main germplasm honeysuckle could be divided by four sets of primers. It provides methodology for improving L. japonica germplasm identification.
关键词(KeyWords):
高分辨率熔解曲线;金银花;种质鉴定
high resolution melting;Lonicerae Japonicae Flos;germplasm identification
基金项目(Foundation): 国家自然科学基金项目(81373959);; 国家科技基础性工作专项(2015FY111500);; 中医药行业科研专项(201407003)
作者(Author):
陈媞颖;蒋超;袁媛;陈康;周骏辉;赵玉洋;黄璐琦;
CHEN Ti-ying;JIANG Chao;YUAN Yuan;CHEN Kang;ZHOU Jun-hui;ZHAO Yu-yang;HUANG Lu-qi;State Key Laboratory of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;
Email:
DOI:
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- 陈媞颖
- 蒋超
- 袁媛
- 陈康
- 周骏辉
- 赵玉洋
- 黄璐琦
CHEN Ti-ying- JIANG Chao
- YUAN Yuan
- CHEN Kang
- ZHOU Jun-hui
- ZHAO Yu-yang
- HUANG Lu-qi
- State Key Laboratory of Dao-di Herbs
- National Resource Center for Chinese Materia Medica
- China Academy of Chinese Medical Sciences
- 陈媞颖
- 蒋超
- 袁媛
- 陈康
- 周骏辉
- 赵玉洋
- 黄璐琦
CHEN Ti-ying- JIANG Chao
- YUAN Yuan
- CHEN Kang
- ZHOU Jun-hui
- ZHAO Yu-yang
- HUANG Lu-qi
- State Key Laboratory of Dao-di Herbs
- National Resource Center for Chinese Materia Medica
- China Academy of Chinese Medical Sciences