油茶SRAP-PCR反应体系的建立与引物筛选

目的]建立油茶SRAP-PCR的反应体系,并筛选合适的引物。方法]采用CTAB法提取油茶基因组DNA,DNA扩增时采用PCR技术,扩增结果采用电泳法和成像系统进行分离和记录。结果]在30μl反应体系中,适宜浓度分别是Mg2＋1.5 mmol/L、模板DNA90ng、引物0.21μmol/L、dNTPs 110μmol/L、TaqDNA聚合酶1.5 U;反应程序中第2次最适退火温度为49℃。随后,在36对引物组合中筛选出11对重复性好、多态性高的引物。结论]优化的SRAP-PCR反应体系及筛选的引物为SRAP分子标记在油茶遗传育种中的应用奠定了基础。

Establishment and Primer Screening of SRAP-PCR System for Camellia oleifera

Objective] The research aimed to establish the SRAP-PCR reaction conditions and select suitable primers.Method]Genomic DNA was isolated from leaves of Camellia oleifera by CTAB method.DNA strip was amplified by PCR technology.The strip was measured and recorded by electrophoresis and imaging system.Result]In 30 μl PCR reaction,the optimum concentration was as follows:Mg2+ of 1.5 mmol/L,template DNA of 90 ng,primer of 0.21 μmol/L,dNTPs of 110 μmol/L,Taq DNA polymerase of 1.5 U.Samples were subjected to thermal profile for amplification of the second annealing temperature at 49℃.11 primer pairs were selected which could amplify stable,polymorphic bands from 36 primer pairs.Conclusion]SRAP-PCR reaction condition and 11 primer pairs provided the basis for genetics and breeding of Camellia oleifera.