【摘要】 目的:发展一种简便快速的制备水稻基因组DNA PCR模板的方法。方法:用枪头捣碎水稻叶片代替液氮研磨法提取水稻基因组DNA作PCR模板,在去污剂SDS和表面活性剂TrionX-100的存在下在沸水中煮沸10min,然后取上清扩增微管蛋白(TubA1)基因内含子。结果:发现在利用煮沸法提取水稻基因组DNA的过程中,用枪头捣碎叶片可代替液氮碾磨,加入0.1%表面活性剂TrionX-100煮沸叶片对制备模板有促进效果,得到了预期的PCR片断。结论:该方法快速、简便、经济,具有良好的重复性与特异性,便于自动化。更多还原

【Abstract】 Objective:To develop a simple method for the rapid preparation of Oryza sativa Indica genomic DNA as PCR template.Method:rice leaf was triturated using pipet tip instead of grounding with liquid nitrogen.Rice leaf was boiled in TE,TE/0.1% SDS and TE/0.1% Triton X-100 respectively and used subsequently for the PCR amplification of an intron of tubulin TubA1 gene .Result:Triturating rice leaves with pipet tips could replace grounding with liquid nitrogen.Subsequent boiling in the presence of 0.1% Triton X-100 for 10 minutes displayed enhancement on PCR amplification.Expected PCR amplicon was obtained with high specificity.Conclusion:This method is simple,rapid,economical,efficient,and apt for automation.更多还原