ZHANG Haokai,DONG Shengjun,LIU Mingguo,et al.Optimization of SSR-PCR Reaction System for Armeniaca sibirica(L.)Lam.[J].Northern Horticulture,2017,41(03):115-120.[doi:10.11937/bfyy.201703027]
山杏SSR-PCR反应体系优化
- Title:
- Optimization of SSR-PCR Reaction System for Armeniaca sibirica(L.)Lam.
- Keywords:
- Armeniaca sibirica(L.)Lam.; SSR-PCR; reaction system; optimization
- 文献标志码:
- A
- 摘要:
- 以4种山杏群体为试材，采用L16（45）正交实验设计，研究了适合4种山杏群体的SSR-PCR反应体系。结果表明：总反应体系为20 μL，其中DNA模板量20 ng，引物浓度0.15 μmol?L-1，Mg2+ 2.0 mmol?L-1，Taq聚合酶量1.0 U，dNTPs 0.25 mmol?L-1。采用引物Y45对43个山杏无性系的DNA进行扩增，扩增产物在100~200 bp，多态性良好，稳定度高，可以用于山杏SSR分子标记研究。
- Abstract:
- Four Armeniaca sibirica(L.)Lam groups were used as test materials，by orthogonal test L16(45)，the optimal SSR-PCR reaction system for Armeniaca sibirica(L.)Lam.was established.The results showed that the optimal systems of SSR（20 μL）was DNA templates 20 ng，primer 0.15 μmol?L-1,Mg2+ 2.0 mmol?L-1,Taq polymerase 1.0 U，dNTPs 0.25 mmol?L-1.After DNA amplification of 43 Armeniaca sibirica(L.)Lam.by using primer Y45，the products were 100-200 bp with good polymorphism and high stability，which could be used for Armeniaca sibirica(L.)Lam.SSR molecular marker development.
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备注/Memo
第一作者简介：张皓凯（1992-），男，辽宁盘锦人，硕士研究生，研究方向为山杏种质资源与多样性。E-mail：zhanghaokai1@126.com.责任作者：董胜君（1974-），男，辽宁瓦房店人，硕士，副教授，现主要从事林木种苗和经济林等研究工作。E-mail:dsj928@163.com.基金项目：中央财政林业科技示范推广资助项目(辽A(2015)01)。