球磨机高通量提取甘蔗叶片DNA研究

High-throughput Genomic DNA Isolation from Sugarcane Leaves using a Mini-Beadbeater

简便快速地获得高质量、高纯度的DNA模板是进行分子遗传学研究和大批量分子检测的基础。常用的液氮手工研磨提取DNA方法存在着耗时、费力、污染等诸多不足。利用从美国引进的球磨机,根据甘蔗叶片的特点,采用国产钢珠,通过优化实验步骤,建立了一种高通量制备甘蔗叶片DNA的方法,其中,叶片快速研磨使用不锈钢珠的直径为4 mm,对叶片进行液氮预冻,时间为30～45 min,叶片用量为50～100 mg,打磨时间为30～40 s。这种方法缩短了提取DNA的时间,提高了效率,而DNA质量与经典方法相当,能够满足分子生物学研究的需要。

It is a primary step to prepare genomic DNA with high quality and purity quickly and economically in study of molecular genetics and mass detection.A common method to extract genomic DNA is to grind sugarcane leaves manually using mortars and pestles in the presence of liquid nitrogen.It is a time-consuming procedure with labors involved and pollution.A fast and efficient protocol to isolate genomic DNA from sugarcane using a mini-beadbeater imported from U.S.A.was established by optimizing pretreatment of sugarcane leaves,types of steel balls in deferent diameters and numbers,grinding time and improvement in experimental procedures and reagents.The improved method was efficient in extracting genomic DNA.In addition,the quality of DNA in this protocol was generally the same in comparison with classic methods,which meets the needs of researches in molecular biology.