Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae

被引:1
|
作者
Alperovich, Nina [1 ]
Scott, Benjamin M. [1 ,2 ,3 ]
Ross, David [1 ]
机构
[1] NIST, Gaithersburg, MD 20899 USA
[2] Univ Maryland, Dept Chem & Biochem, College Pk, MD USA
[3] Univ Saskatchewan, Global Inst Food Secur, Saskatoon, SK, Canada
来源
PLOS ONE | 2023年 / 18卷 / 10期
关键词
RAPID EXTRACTION; YEAST;
D O I
10.1371/journal.pone.0292401
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Although many protocols have been previously developed for genomic DNA (gDNA) extraction from S. cerevisiae, to take advantage of recent advances in laboratory automation and DNA-barcode sequencing, there is a need for automated methods that can provide high-quality gDNA at high efficiency. Here, we describe and demonstrate a fully automated protocol that includes five basic steps: cell wall and RNA digestion, cell lysis, DNA binding to magnetic beads, washing with ethanol, and elution. Our protocol avoids the use of hazardous reagents (e.g., phenol, chloroform), glass beads for mechanical cell disruption, or incubation of samples at 100degree celsius (i.e., boiling). We show that our protocol can extract gDNA with high efficiency both from cells grown in liquid culture and from colonies grown on agar plates. We also show results from gel electrophoresis that demonstrate that the resulting gDNA is of high quality.
引用
收藏
页数:19
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