论文部分内容阅读
DNA提取是SSR-PCR检测的前提,但因棉花中富含多酚、多糖和蛋白质等干扰物质,使得提取棉花DNA较其他植物困难,这也导致棉花分子辅助育种明显滞后.本研究探索出适用于SSR技术的一步快速提取棉花DNA的方法.该方法DNA提取液成份包括0.1 mol/L NaOH、0.7 mmol/L NaC1、20 mmol/L β-巯基乙醇、2％ SDS和2％ PVP.该方法的具体步骤是:将液氮下研磨成粉状的棉花幼嫩叶片,放入含0.5 mL DNA提取液的离心管中.热裂解后加入盐酸离心,取上清,直接用于SSR-PCR扩增.将该方法提取的DNA片段较大,质量较好,能很好地用于TksGlfex DNA聚合酶的SSR-PCR扩增,扩增电泳条带清晰.此棉花DNA提取操作简单、高效,相比传统的棉花DNA提取而言,显著地节约DNA提取时间.“,”DNA extraction is a prerequisite for the detection of SSR-PCR.Because the cotton is rich in polyphenols,polysaccharides and proteins and other interfering substances,which makes the extraction of cotton DNA more difficult than other plants and leads to cotton molecular breeding obviously lagging behind.In this study,a method for rapid extraction of cotton DNA with SSR technology was explored of which DNA extraction buffer include 0.1 mol/L NaOH,0.7 mmol/L NaC1,20 mmol/L β mercaptoethanol,2％ SDS and 2％ PVP.The steps of the method comprise:cotton young leaves grind into powder in liquid nitrogen,put the powder into the tube with 0.5 mL DNA extraction buffer.After thermal cracking,add HCl into the tube,then centrifuge,take supernatant directly for SSR-PCR amplification.The DNA fragment extracted by this method is larger and the quality is better.The DNA can be used for SSR-PCR amplification with TksGlfex polymerase of which the electrophoresis bands are clear.This method of cotton DNA extraction is simple and efficient with only one step centrifugation,which significantly saves DNA extraction time compared to that of the traditional cotton DNA extraction.