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《Acta Agriculturae Zhejiangensis》 2014-01
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Establishment and optimization of AFLP system for Coix

SUN Feng-mei;SHEN Xiao-xia;SHEN Yu-feng;ZHANG Qi-kun;WANG Zhi-an;Zhejiang Chinese Medical University;Zhejiang Research Institute of Traditional Chinese Medicine;Zhejiang Research Institute of Microbilogy;  
By taking 30 samples of Coix species as testing materials,the genetic diversity and genetic relationship was analyzed using AFLP molecular markers technology. The optimal analysis system of AFLP has been established by studying the key factors of conventional process such as gene extraction,restriction digestion and link up,pre-amplication,selective amplification,electrophoresis and siliver staining detection. The result showed that the genome DNA was digested more completely by using two-step method compared with one-step method. In each step,the enzyme reaction time was 4 h added with 15 min for enzyme inactivation. The connection product should be diluted for 10 times and the pre-amplification product should be diluted for 50 times to get a better result. With application of the established system,14 selective amplification primers with better polymorphism were selected out from 64 primers.
【Fund】: 国家中医药管理局中医药行业科研专项(201107011);; 浙江省中药材新品种选育重大科研专项(2012C12912);; 浙江省药用植物种质改良与质量控制技术重点实验室(2011E10015)
【CateGory Index】: S519
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