摘要
介绍了一种以陆地棉不育系104-7A的无菌苗下胚轴为试验材料,专门针对棉花线粒体DNA的提取方法。与黄化苗子叶为材料相比,该法大降低了叶绿体DNA污染的几率。由于棉花中含有大量的棉酚和单宁等物质,增加了线粒体DNA提取的难度,因此在试验的过程中有针对性的加入了PVP、BSA和DIECA,简化了试验流程,提高了线粒体DNA的质量和产率。通过紫外分光光度法、琼脂糖凝胶电泳和PCR检测,证明该方法提取的线粒体DNA纯度极高,能够满足后续试验要求,为研究与线粒体基因相关的工作提供了技术支持。
Using Upland Cotton sterile line 104-7A aseptic seedling hypocotyl as experimental material,this study introduced a kind of method,specifically for cotton mitochondrial DNA extraction.Compared with the etiolated seedling leaves as materials,the method greatly reduced the chloroplast DNA pollution probability.Because Upland Cotton contains large amounts of gossypol and tannin,which made mitochondrial DNA extraction difficult,in the experimental process,we targetly joined PVP,BSA and DIECA,simplified the experimental process and improved the quality and yield of mitochondrial DNA.By ultraviolet spectrophotometry,agarose gel electrophoresis and PCR detection,it proved that high purity of mitochondrial DNA was extracted by this method.The mitochondrial DNA can fully meet the requirements for follow-up experiments.All of these provide technical support for the mitochondrial gene related research.
出处
《华北农学报》
CSCD
北大核心
2012年第B12期61-64,共4页
Acta Agriculturae Boreali-Sinica
基金
国家"863"计划项目(2001AA10A102)
河北省重大技术创新项目(11227118Z)
