高山被孢霉的基因组及转录组的提取技术研究

Technologies of Extracting Genome and Transcriptome from Mortierella alpina

基因组和转录组的高通量测序对样品的纯度和含量要求较高,旨在通过研究高山被孢霉在不同培养条件下和采用不同菌丝处理方法对其基因组和转录组质量的影响来确定最佳提取方式。结果表明,PDA培养基培养的菌丝与发酵培养基培养的菌丝经基因组提取后在琼脂糖凝胶电泳中有明显不同的表现,使用发酵培养基培养的菌丝提取的基因组DNA无降解,而以PDA培养基培养的菌丝提取的基因组DNA出现部分降解。在使用国产试剂盒做高山被孢霉转录组RNA提取实验中发现,使用液氮速冻后破碎菌丝并结合使用β-疏基乙醇更易提取到高浓度且无降解的转录组样品。

High-throughput sequencing of transcriptome and genome demands the high purity and content of sample,the purpose ofthis work is to determine the optimal extraction technology by studying the effects of Mortierella alpina in different culture conditions and usingdifferent hypha processing methods on the genome and transcriptome quality.The results showed that the mycelium from liquid fermentationmedium after genome extraction presented a significant difference in agarose gel electrophoresis compared to the mycelium from PDA culturemedium,the former gave degradation-free genome DNA but the latter was extracted genomic DNA of some degradation.During using domesticbrand kit to study M.alpina tanscriptome RNA extraction,it was found that using liquid nitrogen fast-frozen method to break the hypha cell wallcombined with applying β-Mercaptoethanol resulted in more likely to extract the high concentration and no-degradation transcription sample.