التفاصيل البيبلوغرافية
| العنوان: |
Automation protocol for high-efficiency and high-quality genomic DNA extraction from Saccharomyces cerevisiae. |
| المؤلفون: |
Alperovich, Nina, Scott, Benjamin M., Ross, David |
| المصدر: |
PLoS ONE; 10/17/2023, Vol. 18 Issue 10, p1-19, 19p |
| مصطلحات موضوعية: |
SACCHAROMYCES cerevisiae, DNA, GLASS beads, AUTOMATION, GEL electrophoresis, AGAR, ETHANOL |
| مستخلص: |
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. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
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