DNA isolation / purification Yeast - Pichia pastoris

DNA isolation and purification is the first critical step in sample preparation that helps ensure optimal performance of downstream assays like PCR, microarrays, and sequencing. Failure in yielding high-quality DNA would be the major reason that DNA doesn't work for the downstream application. To circumvent this, one should follow the recommended storage conditions to minimize DNA degradation by nucleases and shouldn't overload the purification system.

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4 years ago

4 years ago by Jean Rossignol France

How to avoid genomic DNA contamination when isolating mitochondrial DNA?

Greetings! I am trying to isolate mitochondrial DNA from S. cerevisiae while avoiding contamination from genomic DNA. Any and all help is greatly appreciated.

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Protocol tips
Susceptibility to yeast lytic enzymes varies for different yeast

If you see incomplete lysis, extend the first incubation time up to 2 hours or over 16 hours.
Protocol tips
For blood DNA isolation, blood must be collected in EDTA, heparin or citrate anticoagulant tubes to prevent clotting.
Downstream tips
- Include RNAse treatment for 15-20 min.
- Use prewarmed TE buffer to elute the DNA
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