摘要
为建立最佳的宫粉紫荆SRAP-PCR反应体系,采用单因素和L16(45)正交试验设计对反应体系中的模板DNA、Mg2+、引物浓度、d NTPs和Taq聚合酶进行优化。表明宫粉紫荆SRAP-PCR 25μL反应体系的最佳组合为:模板DNA 50 ng、Mg2+2.25 mmol/L、引物0.25μmol/L、d NTPs 0.30 mmol/L、Taq酶1.5 U。并利用优化的SRAP-PCR体系进行验证,表明不同的宫粉紫荆样本均能扩增出清晰且带型基本一致的谱带,表明本试验建立的SRAP-PCR体系稳定,可用于今后开展宫粉紫荆种质资源遗传多样性研究、品种鉴定、优良品种筛选和近缘种杂交育种等研究工作。
In order to establ ish the optimal SRAP system for Bauhinia variegata Linn., the concentration of template DNA, Mg2＋, primers, d NTPs and Taq polymerase were determine their optimal levels by using singlefactor and the orthogonal experimental design L16（45）. The results showed that the optimal SRAP-PCR system for Bauhinia variegata Linn. was established on the basis of orthogonal experiment,which was 25 μL capacity of reaction system contained 50 ng template DNA, 2.25 mmol/L Mg2＋, 0.25 μmol/L primers, 0.30 mmol/L d NTPs and 1.5 U Taq polymerase. This optimized system was verified by different samples of Bauhinia variegata Linn.,and primer pairs were screened out via their stable amplification and clear banding patterns. The results showed that the SRAP-PCR system was stable and reliable and it would lay the foundation for genetic diversity,germplasm identification, selection of elite cultivars and crossbreeding of related genus.
出处
《分子植物育种》
CAS
CSCD
北大核心
2017年第10期3989-3996,共8页
Molecular Plant Breeding
基金
广州市科技计划项目(201300000105)
越秀区科技计划项目(2015-CY-008)
2014年度生态园林产业技术创新战略联盟自设科研课题共同资助
关键词
宫粉紫荆
SRAP
体系优化
正交试验设计
Bauhinia variegata Linn., SRAP, System optimization, Orthogonal experimental design
