适合于全长cDNA文库构建的猪苓菌核及菌丝体总RNA提取方法比较

目的]筛选出适合于c DNA文库构建的猪苓菌核及菌丝体总RNA的提取方法。方法]采用Trizol改良法和EASY Spin植物快速提取试剂盒法(简称试剂盒法)对猪苓菌丝及菌核的总RNA进行提取。结果]经紫外分光光度计对所得总RNA的均一性及1%琼脂糖凝胶电泳对所得总RNA的完整性检测后发现,2种方法均可提出完整的总RNA,试剂盒法相较于Trizol改良法其所提取的总RNA质量和纯度都较好,但Trizol改良法所提取的总RNA产率比较高。结论]LD-PCR成功合成了猪苓菌核和菌丝体的c DNA产物,呈弥散状分布在1 0005 000 bp和7502 500 bp,满足c DNA建库的要求。

Comparison of Methods for Extracting Total RNA from Polyporus umbellatus Mycelium and Sclerotium Suitable for Constructing Full-length cDNA Library

Objective] To screen out the method for extracting total RNA from Polyporus umbellatus mycelium and sclerotium suitable for cDNA library construction.Method]The total RNA of mycelium and sclerotium of P.umbellatus was extracted by improved Trizol method and EASY Spin plant RNA kit method(kit method for short).Result]The total RNA was homogenized by UV spectrophotometer and the integrity of the obtained total RNA was detected by 1％ agarose gel electrophoresis.It was found that both methods could provide the complete total RNA.Compared with the kit method,the total RNA extracted by the improved Trizol method was better in quality and purity,but the total RNA extracted by improved Trizol method had higher yield.The cDNA of mycelium and sclerotium of P.umbellatus was successfully synthesized by LD-PCR and distributed in the range of 1 000-5 000 bp and 750-2 500 bp,which satisfied the requirement of cDNA library construction.