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Position: Home > Articles > Establishment and Optimization of Sequence-related Amplified Polymorphism Amplification System for Rehmannia glutinosa Hubei Agricultural Sciences 2009,48 (3) 33-37

怀地黄SRAP分子标记优化体系的建立

作  者:
周春娥;谷凤平;路淑霞;段红英;周延清
单  位:
河南师范大学生命科学学院
关键词:
怀地黄;相关序列扩增多态性分子标记;扩增体系优化
摘  要:
为了建立适宜怀地黄的SRAP反应体系,以22个不同类型的怀地黄品种为材料,研究了PCR反应体系的主要成分对SRAP扩增结果的影响。对SRAP反应体系中的DNA模板浓度、TaqDNA聚合酶浓度、Mg2+浓度、引物浓度以及dNTP浓度进行了探索,确立的适合怀地黄SRAP反应的体系为:在25μL的反应体系中,模板DNA20ng、2.5mmol·L-1Mg2+、0.32μmol·L-1的上下游引物、0.30mmol·L-1的dNTPs以及2.5UTaq酶。并利用该反应体系对怀地黄22个不同品种进行了SRAP反应,发现不同品种间的DNA谱带多态性丰富,证实该体系稳定可靠,可以用于怀地黄的分子标记研究。
译  名:
Establishment and Optimization of Sequence-related Amplified Polymorphism Amplification System for Rehmannia glutinosa
作  者:
ZHOU Chun-e,GU Feng-ping,LU Shu-xia,DUAN Hong-ying,ZHOU Yan-qing (College of Life Science, Henan Normal University, Xinxiang 453007,Henan,China)
关键词:
Rehmannia glutinosa L.; sequence-related amplified polymorphism; optimization of amplification protocol
摘  要:
In this study, SRAP-PCR amplified condition was optimized on 22 Rehmannia glutinosa lines and analyzed the effects of the concentration of DNA template,Taq DNA polymerase, primer Mg2+, dNTP on PCR results, The results showed that: 2.5 mmol·L-1 Mg2+, 0.30 mmol·L-1 dNTP mixture, 2.5U Taq DNA polymerase, 0.32 μmol·L-1 each primer, 20 ng template DNA and 2.5 μL 10×PCR buffer in 25 μL SRAP reaction system were the best suitable PCR system. SRAP of 22 different R.glutinosa lines were obtanied and detected. Polymorphism between different lines abundantly detected by 2% agarose gel which showed this system was suitable and stable.

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