摘要
该文筛选一种适用于黑曲霉全基因组测序的DNA提取方法。通过对黑曲霉菌体培养条件、细胞壁破裂方法和常见DNA提取方法的比较研究,测定DNA纯度、条带完整性,获得一套黑曲霉全基因组DNA提取方法。结果表明,菌丝体经液氮充分研磨,采用CTAB法进行基因组DNA提取,并结合试剂盒纯化法,适用于黑曲霉全基因组测序的DNA提取方法。该方法提取不同分离源的35株黑曲霉基因组DNA,检测结果蛋白质和RNA含量低,均有主条带,浓度为52.24~212.37 ng/μL,满足全基因组测序要求。该研究筛选的黑曲霉全基因组DNA提取法可以满足分析要求,具有在实验室推广应用价值。
To screen a method of DNAextraction suitable for whole genome sequencing of Aspergillus niger,the culture conditions of A.niger,cell wall rupture methods and common DNA extraction methods were compared and the DNA purity and band integrity were determined.The results showed that mycelium fully milled by liquid nitrogen,genomic DNA extracted by CTAB method,and combining with the kit purification was a DNA extraction method suitable for sequencing the whole genome of A.niger.Using this method to extract the genomic DNA of 35 strains of A.niger isolated from different sources,the results showed that the content of protein and RNA was low,and the main bands were bright and clear,with the concentrations ranging from 52.24 to 212.37 ng/μL,which met the requirements of whole-genome sequencing.The whole genome DNA extraction method of A.niger screened in this study can meet the analysis requirements and has the value of popularization and application in laboratory.
作者
蔡程山
王雨
白飞荣
翟磊
张天赐
胡海蓉
姚粟
CAI Chengshan;WANG Yu;BAI Feirong;ZHAI Lei;ZHANG Tianci;HU Hairong;YAO Su(China Center of Industrial CultureCollection,China National Research Institute of Food and Fermentation Industries Co.Ltd.,Beijing 100015,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2020年第6期13-18,共6页
Food and Fermentation Industries
基金
国家重点研发计划资助(2018YFC1603800)。
