LIU Meng-ya,HUANG Ru-kui,HUANG Yu-hui,et al.Orthogonal Design-direct Analysis and SSR for SRAP Amplification System in Momordica charantia[J].Northern Horticulture,2014,38(05):85-88.
正交直观分析法和新复极差法优化苦瓜SRAP反应体系研究
- Title:
- Orthogonal Design-direct Analysis and SSR for SRAP Amplification System in Momordica charantia
- 文章编号:
- 1001-0009(2014)05-0085-04
- Keywords:
- Momordica charantia; orthogonal design-direct analysis; SSR; SRAP
- 分类号:
- S 642.5
- 文献标志码:
- A
- 摘要:
- 以苦瓜为试材,采用正交直观分析法和新复极差法相结合的方法,对影响苦瓜SRAP 反应体系的5种因素(dNTP浓度、模板DNA、引物浓度、Mg2+浓度及Taq DNA聚合酶)4个水平进行优化筛选,以期优化苦瓜SRAP的PCR反应体系。结果表明:苦瓜SRAP分析的优化反应体系为20 μL PCR 反应体系中含有1× PCR buffer,250 μmol/L dNTP,50 ng模板DNA,1.2 μmol/L引物,1.5 mmol/L Mg2+,1.5 U Taq DNA聚合酶。
- Abstract:
- Taking Momordica charantia as test material,using SSR and orthogonal design-direct analysis to optimize the four levels of five factor(dNTP,template DNA,primer,Mg2+,Taq DNA polymerase).The results showed that the optimum SRAP-PCR reaction system contains 1×PCR buffer,250 μmoL/L dNTP,50 ng template DNA,1.2 μmoL/L primer,1.5 mmoL/L Mg2+,1.5 U Taq DNA polymerase,in a total volume of 20 μL SRAP-PCR system.
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备注/Memo
第一作者简介:刘萌芽(1989-),女,河南洛阳人,硕士研究生,现主要从事苦瓜抗病性鉴定工作。E-mail:mengya0316@126.com.