26S rDNA D1/D2区域序列同源性分析是一种常用的酵母分子鉴定方法。D1/D2区域序列的扩增需要酵母染色体DNA作为模板,目前常用的酵母染色体DNA提取方法繁琐耗时且难以进行批量操作,限制了酵母分子鉴定的规模化。研究中建立了一种快速高效的批量酵母染色体DNA提取方法,整个提取过程仅耗时20min,从而使得酵母大规模快速分子鉴定成为可能。用该方法制备的染色体DNA不需要任何后续处理即可作为模板用于扩增酵母的26S rDNA D1/D2区域序列,获得了良好的扩增结果以及扩增产物的测序结果。

D1/D2 domain of 26S rDNA sequence-based alignment and analysis is a widely used method for yeast identification and classification.Rapid and repeatable protocols to extract and prepare the chromosomal DNA of yeast are the basic needs to acquire the templates for following amplification of D1/D2 domain.However,the present methods to extract chromosomal DNA are time-consuming and unable to deal with large-scale samples at the same time.In the present study,a novel rapid and efficient method to identify a large amount of yeast isolates in the same batch was established.The chromosomal DNA required only 20 minutes to prepare and can be directly used as template for PCR amplification.Results of amplified ITS sequences were clear and easy to identify by sequencing analysis.