摘要
为建立可靠并且稳定的SRAP-PCR反应体系,进一步开展新疆薰衣草种质资源的遗传多样性研究提供帮助。本研究采用L16(45)正交试验设计方法,对薰衣草SRAP-PCR反应体系主要影响因素DNA模板、Mg^2+、dNTPs、引物浓度及Taq DNA聚合酶的用量进行了筛选,比较了5个因素对扩增效果的影响。试验结果表明,各因素水平变化对反应体系的影响从大到小依次为:Mg^2+>引物>Tap聚合酶>dNTPs>DNA;建立的最佳反应体系为:总体积25.0μL,2.5μL 10×Buffer、Mg^2+浓度3.0 mmol/L、d NTPs浓度0.32 mmol/L、引物浓度0.24μmol/L、DNA模板量60 ng、Taq DNA聚合酶用量0.4 U,该体系经验证可以扩增出清晰、稳定且多态性较好的条带,可为开展后续的研究提供技术基础。
In order to establish a reliable and stable SRAP-PCR reaction system,and to further study the genetic diversity of lavender germplasm resources in Xinjiang.In this study,L16(45)orthogonal experimental design method was used to optimize the main influencing factors of lavender SRAP-PCR reaction system DNA template,Mg^2+,d NTPs,primer concentration and Taq DNA polymerase,and compared five factors to expand effect of increasing effect.The reaction system is consisted by the following components:2.5μL 10×Buffer,3.0 mmol/L Mg2^+,0.32 mmol/L d NTPs,0.24μmol/L primers,60 ng DNA and 0.4 U DNA polymerase.The influence of the level of each factor on the reaction system is ranked as follows:Mg2+>primer>Taq DNA polymerase>dNTPs>DNA.The system has been proved to be able to amplify clear,stable and polymorphism bands,which can provide a technical basis for further research.
作者
阿迪莱·阿布都热依木
苏秀娟
吕雪梅
周界光
尹松松
龚林涛
曲延英
Adilai Abudoureyimu;Su Xiujuan;Lu Xuemei;Zhou Jieguang;Yin Songsong;Gong Lintao;Qu Yanying(College of Agriculture,Xinjiang Agricultural University,Urumqi,830052)
出处
《分子植物育种》
CAS
CSCD
北大核心
2020年第19期6418-6423,共6页
Molecular Plant Breeding
基金
新疆农业大学作物学重点学科发展基金项目(XNCDKY2018010)
国家自然科学基金(31460383)
新疆维吾尔自治区高校科研计划科学研究重点项目(XJEDU2014I069)
新疆维吾尔自治区自然科学基金(2016D01A035)
新疆维吾尔自治区重点研发项目(2017B03016-1)共同资助。
