摘要
介绍了一种在提取小麦DNA实验中常用的CTAB提取方法基础上改良的微量DNA提取方法。通过提取多个材料的DNA并经分光光度计检测,琼脂精电泳结果表明,这种方法提取的DNAOD260/280在2．00- 1．8间波动,较常规大量法提取DNA的质量高,能满足100-200余次的PCR反应的需要。采用该法对提取的普通小麦DNA以及转抗除草剂基因Bar-的转基因值株DNA分别进行SSR标记检测和PCR扩增,结果表明 SSR标记扩增效果与大量法无明显的差异,抗性植株DNA能扩增出Bar基因的特征带。
This article introduced a modified micromethod of isolating wheat genomic DNA based on the traditional CTAB method. The agarose electrophoresis suggested that the higher-quality DNA obtained by this micromethod(OD260/280 = 2. 00- 1. 8), Meanwhile, about one hundred to two hundred PCR reactions could be met. Then, this method have been applied for isolating DNA of transgenic wheat and common wheat (non-transgenic) to amplified the given fragment of the Bar gene and SSR markers identification, respectively. The results indicated the 310 bp target fragment was amplified on the transgenic wheat, and the similarity SSR amplifying results was detected between the DNA isolated by the micromethod and traditional CTAB method.
出处
《西北农业学报》
CAS
CSCD
北大核心
2005年第6期83-86,共4页
Acta Agriculturae Boreali-occidentalis Sinica
基金
四川省科技厅应用基础项目(2002A008)四川省教育厅重点项目(03JY029-028-2863)国家"863"项目(2003AA207100)资助
