摘要
改良发展一种快速简便、高效的小麦基因组DNA提取方法,命名为TENR提取法。以春小麦Thatcher和以Thatcher为遗传背景的近等基因系TcLr19、TcLr20、TcLr28为试验材料,采用TENR提取法提取其基因组DNA,DNA提取质量通过紫外分光光度分析法和琼脂糖凝胶电泳法并结合抗叶锈基因Lr20的STS标记、Lr19和Lr28的SCAR标记、Lr28的SSR标记进行PCR扩增检测进行评价。结果表明,TENR提取法获得的DNA质量合格,且能扩增出条带清晰正确的分子标记目的片段。该方法能够获得适用于STS、SCAR、SSR标记PCR扩增的高质量的DNA,而且加快了DNA提取速度、降低了成本、减少了污染源,为大批量提取DNA提供了技术支撑。
One easy and efficient wheat genomic DNA extraction method had been developed and named as TENR extraction method. Spring wheat cultivar Thatcher, the near-isogenic lines TcLr19, TcLr20 and TcLr28 which taken the Thatcher as the genetic background were used as the materials and the DNA were extracted by TENR method. The quality of DNA were evaluated by UV spectrophotometric analysis, agarose gel electrophoresis method and PCR amplification using the STS marker for leaf rust resistance gene Lr20, SCAR markers for Lr19 and Lr28, SSR marker for Lr28. The results showed that high qualified DNA was obtained by TENR extraction method, clear and right bands were amplified in the test. The method could reduce the DNA extracting processing, costs, pollution to the environment, and get high quality DNA for PCR analysis. It provided base for the preparation of large number of samples for PCR-based assays.
出处
《中国农学通报》
CSCD
北大核心
2010年第19期22-26,共5页
Chinese Agricultural Science Bulletin
基金
国家公益性行业(农业)科研专项(200903035)
国家"十一五"支撑计划(2006BAD08A05)
