Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae.
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...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2023-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0292401&type=printable |
| _version_ | 1797649547756830720 |
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| author | Nina Alperovich Benjamin M Scott David Ross |
| author_facet | Nina Alperovich Benjamin M Scott David Ross |
| author_sort | Nina Alperovich |
| collection | DOAJ |
| description | 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 100°C (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. |
| first_indexed | 2024-03-11T15:48:39Z |
| format | Article |
| id | doaj.art-0c19becc441c419ea7a7591b3d2ffcb3 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-03-11T15:48:39Z |
| publishDate | 2023-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-0c19becc441c419ea7a7591b3d2ffcb32023-10-26T05:31:47ZengPublic Library of Science (PLoS)PLoS ONE1932-62032023-01-011810e029240110.1371/journal.pone.0292401Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae.Nina AlperovichBenjamin M ScottDavid RossAlthough 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 100°C (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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0292401&type=printable |
| spellingShingle | Nina Alperovich Benjamin M Scott David Ross Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae. PLoS ONE |
| title | Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae. |
| title_full | Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae. |
| title_fullStr | Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae. |
| title_full_unstemmed | Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae. |
| title_short | Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae. |
| title_sort | automation protocol for high efficiency and high quality genomic dna extraction from saccharomyces cerevisiae |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0292401&type=printable |
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