ZHANG Guo-wu,HUANG Tao,WU Nan-sheng,et al.Establishment and Optimization of ISSR-PCR Reaction System for Polyscias[J].Northern Horticulture,2014,38(15):101-105.
观叶福禄桐ISSR-PCR反应体系的建立及优化
- Title:
- Establishment and Optimization of ISSR-PCR Reaction System for Polyscias
- 文章编号:
- 1001-0009(2014)15-0101-05
- 关键词:
- 福禄桐; ISSR-PCR反应; 梯度PCR; 退火温度
- Keywords:
- .Polyscias; ISSR-PCR reaction; gradient PCR; annealing temperature
- 分类号:
- S682.36
- 文献标志码:
- A
- 摘要:
- 以福禄桐叶片为试材,采用正交实验和单因子试验,分析模板DNA、Mg2+、Taq酶、引物和dNTP五个因子对福禄桐ISSR-PCR反应的影响。结果表明:福禄桐ISSR-PCR最佳反应体系为在25 μL反应体系中,模板DNA为15 ng,Mg2+3.0 mmol/L,Taq酶0.5 U,引物0.6 μmol/L, dNTP 0.20 mmol/L,通过梯度PCR试验得到相应引物最佳退火温度为47℃;该试验可为福禄桐遗传多样性分析和亲缘关系等提供理论基础。
- Abstract:
- Use leaves of Polyscias as materials,the effect of the five main reaction elements (Mg2+,Taq DNA polymerase,dNTP,DNA template and primer) on Polyscias ISSR-PCR were all optimized by combining single factor experiments and orthogonal test method.The results showed that the best ISSRPCR reaction system of Polyscias was 25 μL,reaction system concention:the DNA template 15.00 ng,Mg2+3.0 mmol/L,Taq DNA polymerase 0.5 U,primer 0.6 μmol/L,dNTP 0.20 mmol/L.The optimal annealing temperature of ISSR-PCR was 47℃ by gradient PCR.This system could provide theoretical foundation for carrot research on genetic diversity analysis,genetic relationship,etc
参考文献/References:
[1]李克烈,王荣香,陈伟,等.羽叶南洋参的组织培养[J].植物生理学通讯,2007,43(2):324-325.
相似文献/References:
[1]赵雷,张继兰.云南松ISSR-PCR反应体系建立及优化[J].北方园艺,2012,36(12):130.
ZHAOLei,ZHANGJi-lan.EstablishmentandOptimizationofISSR-PCRReactionSystemfor PinusyunnanensisFranch[J].Northern Horticulture,2012,36(15):130.
备注/Memo
第一作者简介:张国武(1966-),男,博士,高级工程师,研究方向为森林培育。