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

被引:2
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
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页数:19
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共 31 条
  • [1] Genome-scale engineering of Saccharomyces cerevisiae with single-nucleotide precision
    Bao, Zehua
    HamediRad, Mohammad
    Xue, Pu
    Xiao, Han
    Tasan, Ipek
    Chao, Ran
    Liang, Jing
    Zhao, Huimin
    [J]. NATURE BIOTECHNOLOGY, 2018, 36 (06) : 505 - +
  • [2] Rapid host strain improvement by in vivo rearrangement of a synthetic yeast chromosome
    Blount, B. A.
    Gowers, G-O. F.
    Ho, J. C. H.
    Ledesma-Amaro, R.
    Jovicevic, D.
    McKiernan, R. M.
    Xie, Z. X.
    Li, B. Z.
    Yuan, Y. J.
    Ellis, T.
    [J]. NATURE COMMUNICATIONS, 2018, 9
  • [3] GC Preps: Fast and Easy Extraction of Stable Yeast Genomic DNA
    Blount, Benjamin A.
    Driessen, Maureen R. M.
    Ellis, Tom
    [J]. SCIENTIFIC REPORTS, 2016, 6
  • [4] Extremely rapid extraction of DNA from bacteria and yeasts
    Cheng, HR
    Jiang, N
    [J]. BIOTECHNOLOGY LETTERS, 2006, 28 (01) : 55 - 59
  • [5] Clikeman JA, 2001, GENETICS, V157, P579
  • [6] Engineered yeast genomes accurately assembled from pure and mixed samples
    Collins, Joseph H.
    Keating, Kevin W.
    Jones, Trent R.
    Balaji, Shravani
    Marsan, Celeste B.
    Como, Marina
    Newlon, Zachary J.
    Mitchell, Tom
    Bartley, Bryan
    Adler, Aaron
    Roehner, Nicholas
    Young, Eric M.
    [J]. NATURE COMMUNICATIONS, 2021, 12 (01)
  • [7] Rapid isolation of yeast genomic DNA: Bust n' Grab
    Harju, S
    Fedosyuk, H
    Peterson, KR
    [J]. BMC BIOTECHNOLOGY, 2004, 4 (1)
  • [8] Precise control of SCRaMbLE in synthetic haploid and diploid yeast
    Jia, Bin
    Wu, Yi
    Li, Bing-Zhi
    Mitchell, Leslie A.
    Liu, Hong
    Pan, Shuo
    Wang, Juan
    Zhang, Hao-Ran
    Jia, Nan
    Li, Bo
    Shen, Michael
    Xie, Ze-Xiong
    Liu, Duo
    Cao, Ying-Xiu
    Li, Xia
    Zhou, Xiao
    Qi, Hao
    Boeke, Jef D.
    Yuan, Ying-Jin
    [J]. NATURE COMMUNICATIONS, 2018, 9
  • [9] Koetsier GC., 2019, New England Biolabs Inc
  • [10] Synthetic Genomics From a Yeast Perspective
    Koster, Charlotte C.
    Postma, Eline D.
    Knibbe, Ewout
    Cleij, Celine
    Daran-Lapujade, Pascale
    [J]. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, 2022, 10