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
相关论文
共 50 条
  • [1] A protocol for high-quality genomic DNA extraction from legumes
    Agbagwa, I. O.
    Datta, S.
    Patil, P. G.
    Singh, P.
    Nadarajan, N.
    GENETICS AND MOLECULAR RESEARCH, 2012, 11 (04) : 4632 - 4639
  • [2] A high-throughput, high-quality plant genomic DNA extraction protocol
    Li, H.
    Li, J.
    Cong, X. H.
    Duan, Y. B.
    Li, L.
    Wei, P. C.
    Lu, X. Z.
    Yang, J. B.
    GENETICS AND MOLECULAR RESEARCH, 2013, 12 (04): : 4526 - 4539
  • [3] High-Quality Genomic DNA Extraction Protocol for Bacillus and Clostridium Species
    Mekonnen, Yohannes Beyene
    Aspholm, Marina Elisabeth
    Zegeye, Ephrem Debebe
    CURRENT PROTOCOLS, 2024, 4 (10):
  • [4] EXTRACTION OF HIGH-QUALITY GENOMIC DNA FROM MICROSAMPLES OF HUMAN BLOOD
    MA, HW
    CHENG, J
    CADDY, B
    JOURNAL OF THE FORENSIC SCIENCE SOCIETY, 1994, 34 (04): : 231 - 236
  • [5] Extraction of high-quality genomic DNA from latex-containing plants
    Michiels, A
    Van den Ende, W
    Tucker, M
    Van Riet, L
    Van Laere, A
    ANALYTICAL BIOCHEMISTRY, 2003, 315 (01) : 85 - 89
  • [6] An inexpensive, automation-friendly protocol for recovering high-quality DNA
    Ivanova, Natalia V.
    Dewaard, Jeremy R.
    Hebert, Paul D. N.
    MOLECULAR ECOLOGY NOTES, 2006, 6 (04): : 998 - 1002
  • [7] A Protocol for Extraction of High-quality RNA and DNA from Peanut Plant Tissues
    Dongmei Yin
    Haiying Liu
    Xingguo Zhang
    Dangqun Cui
    Molecular Biotechnology, 2011, 49 : 187 - 191
  • [8] A Protocol for Extraction of High-quality RNA and DNA from Peanut Plant Tissues
    Yin, Dongmei
    Liu, Haiying
    Zhang, Xingguo
    Cui, Dangqun
    MOLECULAR BIOTECHNOLOGY, 2011, 49 (02) : 187 - 191
  • [9] HIGH-EFFICIENCY TRANSFORMATION OF SACCHAROMYCES-CEREVISIAE BY ELECTROPORATION
    MANIVASAKAM, P
    SCHIESTL, RH
    NUCLEIC ACIDS RESEARCH, 1993, 21 (18) : 4414 - 4415
  • [10] Optimized Protocol for Simple Extraction of High-Quality Genomic DNA from Clostridium difficile for Whole-Genome Sequencing
    Sim, James Heng Chiak
    Anikst, Victoria
    Lohith, Akshar
    Pourmand, Nader
    Banaei, Niaz
    JOURNAL OF CLINICAL MICROBIOLOGY, 2015, 53 (07) : 2329 - 2331