Thermo Scientific Pierce Yeast DNA Extraction Reagent Kit utilizes Thermo Scientific Y-PER Reagent to quickly and easily lyse yeast cells to release DNA for study.
This kit provides greater yields than traditional methods of DNA isolation from yeast. Typically, extraction and purification of DNA from yeast are time-consuming and labor-intensive. The yeast proteinaceous cell wall is notoriously difficult to lyse and requires harsh treatments that are time-consuming and can damage the extracted DNA. The Yeast DNA Extraction Kit protocol requires less than one hour to complete, is effective without enzymatic treatment or glass beads, and yields little to no RNA contamination regardless of RNase presence. In studies with Saccharomyces cerevisiae, high yields of genomic and plasmid DNA are consistently obtained. DNA purified using this kit is suitable for PCR amplification, bacterial transformations, restriction digestions and hybridization applications.
Highlights:
- Eliminates need for glass beads or harsh enzyme treatments
- Yields DNA that is compatible with polymerase chain reaction (PCR) amplification
- Rapidly isolates plasmid DNA from S. cerevisiae suitable for transformation of E. coli
- Protocol is easily scalable for use with single colonies or 500mL cultures
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Extraction of yeast genomic DNA and subsequent PCR amplification. Thermo Scientific Yeast DNA Extraction Kit was used to extract and purify DNA from S. cerevisiae strain DY150 that had been transformed with a plasmid harboring the gene for green fluorescent protein (GFP). The purified DNA was used to amplify the chromosomal ACT1 and UME6 genes and the gene-encoding GFP carried on the plasmid.
Lane 1. Lambda DNA digested with Hind III;
Lane 2. S. cerevisiae genomic DNA (small band, 5 kbp, corresponds to dsRNA from the yeast killer virus);
Lane 3. PCR amplification of GFP from plasmid;
Lane 4. PCR amplification of chromosomal ACT1 gene, and Lane 5. PCR amplification of chromosomal UME6 gene.
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