食品与发酵工业 >

2020 , Vol. 46 >Issue 6: 13 - 18

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.022988

研究报告

黑曲霉Aspergillus niger全基因组DNA提取方法的改良与比较

  • 蔡程山 ,
  • 王雨 ,
  • 白飞荣 ,
  • 翟磊 ,
  • 张天赐 ,
  • 胡海蓉 ,
  • 姚粟
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  • (中国食品发酵工业研究院有限公司, 中国工业微生物菌种保藏中心,北京,100015)
硕士,助理工程师(姚粟教授级高级工程师为通讯作者, E-mail:milly@china-cicc.org)

收稿日期: 2019-12-06

  网络出版日期: 2020-04-24

基金资助

国家重点研发计划资助(2018YFC1603800)

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Improvement and comparison of Aspergillus niger genomic DNA extraction methods

  • CAI Chengshan ,
  • WANG Yu ,
  • BAI Feirong ,
  • ZHAI Lei ,
  • ZHANG Tianci ,
  • HU Hairong ,
  • YAO Su
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  • (China Center of Industrial CultureCollection, China National Research Institute of Food and Fermentation Industries Co. Ltd., Beijing 100015, China)

Received date: 2019-12-06

  Online published: 2020-04-24

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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。

关键词： 黑曲霉; DNA提取; 全基因组测序; DNA完整性

本文引用格式

蔡程山 , 王雨 , 白飞荣 , 翟磊 , 张天赐 , 胡海蓉 , 姚粟 . 黑曲霉Aspergillus niger全基因组DNA提取方法的改良与比较[J]. 食品与发酵工业, 2020 , 46(6) : 13 -18 . DOI: 10.13995/j.cnki.11-1802/ts.022988

Abstract

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.

Key words： Aspergillus niger; DNA extraction; whole genome sequencing; DNA integrity

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