Abstract
In this protocol, yeast DNA is prepared by digestion of the cell wall and lysis of the resulting spheroplasts with SDS. This method reproducibly yields several micrograms of yeast DNA that can be efficiently cleaved by restriction enzymes and used as a template in polymerase chain reaction (PCR). Note that yeast colonies can also be used directly in PCR, without purifying yeast DNA.
© 2020 Cold Spring Harbor Laboratory Press.
MeSH terms
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Cell Wall / chemistry*
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Cloning, Molecular / methods
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Culture Media / chemistry
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DNA Restriction Enzymes / metabolism
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DNA, Fungal / genetics*
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DNA, Fungal / isolation & purification
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DNA, Fungal / metabolism
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Genomic Library
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Polymerase Chain Reaction / methods*
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / growth & development
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Sodium Dodecyl Sulfate / chemistry*
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Spheroplasts / chemistry*
Substances
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Culture Media
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DNA, Fungal
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Sodium Dodecyl Sulfate
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DNA Restriction Enzymes