大麦高分子量基因组DNA提取的优化

高分子量(HMW)DNA的制备是进行生物基因组物理作图等相关基因组学研究至关重要的基础工作。为建立一种高效的提取大麦高分子量DNA的方法,对琼脂糖包埋原生质体和琼脂糖包埋细胞核分离基因组DNA的方法进行了改良和优化。限制酶消化,脉冲电场凝胶电泳(PFGE)和Southern杂交等检测表明,应用优化的实验方法所分离的大麦基因组HMW DNA质量好,数量多。与原方法相比,新改进的方法实验流程缩短,效率提高,所分离HMWDNA能更好地满足后续相关研究的需要。这两种优化的分离大麦HMW DNA技术亦可应用于其它禾谷类植物基因组DNA的制备。

Optimization of High Molecular Weight DNA Isolation from Hordeum vulgare Genome

The production and preparation of high molecular weight(HMW) DNA is an essential basic work for such research of related genomics as physical mapping of biological genome.In order to build an effective extraction method of Hordeum vulgare HMW DNA,the study improved and optimized the common methods of embedding protolasts or nuclei in agarose plugs to isolate Hordeum vulgare genomic DNA.The HMW DNA prepared from the optimized methods was identified by the restriction enzyme digestion,pulsed-field gel electrophoresis separation and Southern hybridization.The results showed that the optimized methods gave higher quantity and better quality of HMW DNA.In comparison with the original methods,the improved methods had the shorter experiment process and higher efficiency.The HMW DNA from the improved methods can better satisfy the follow-up studies.The technique of optimized methods of isolating can also be applied to HMW DNA isolation from other cereal genome.