[关键词]
[摘要]
从美国NCBI数据库中下载了286 868条亚麻EST,利用SSRIT软件检索出符合要求的SSR重复序列4 310条,占整个EST数据库的1.50%。其中三核苷酸出现频率最高,占总SSR的40.09%;SSR重复类型总共235种,序列中占总SSR比例大于1%的重复类型共有21种,占SSR序列总数的68.63%。通过正交设计(L16)得到了胡麻EST-SSR最佳PCR反应体系:即20 μL 反应体系里,引物0.2 μmol/L、模板DNA 50 ng、dNTP 0.10 mmol/L、Mg2+ 1.5 mmol/L、Taq DNA聚合酶0.50 U/μl、2 μL 10×buffer,其余用ddH2O 补齐。并通过单因素优化试验,对得到的胡麻EST-SSR最佳PCR反应体系进行了验证,结果完全一致。
[Key word]
[Abstract]
In this study, 286 868 expressed sequence tags(ESTs) are downloaded from the USA NCBI database. 4 310 appropriate simple sequence repeats(SSRs) are picked out with software of SSRIT. These SSRs accounted for 1.50% of the EST database and 40.09% of them is trinucleotide which is the maximum type. A total of 235 SSR types are got, 21 of them accounted for 68.63% total SSRs picked out in this study, and each of the 21 SSRs is more than 1% of the total SSRs. The optimal PCR reaction system for EST-SSR of flax is got by orthogonal design as follow: primer 0.2 μmol/L; template DNA, 50 ng, dNTP, 0.10 mmol/L、 Mg2+ , 1.5 mmol/L、 Taq DNA polymerase 0.50 U/μl、 2 μL 10×buffer, and at last, filling 20 μL with ddH2O. The consistent result is got from single factor optimization experiment on he optimal PCR reaction system for EST-SSR of flax.
[中图分类号]
S565.9
[基金项目]
国家自然基金项目(31160291);国家自然基金项目(31560347);兰州市人才创新创业科技计划(2014-rc-44)部分内容。