Dear all,
I am looking for recommendations for an efficient protocol for the extraction of Genomic DNA from several fungal cultures.
I have most of the chemicals and equipment used in the process, but I am not getting a complete procedure (Starting from Cell disruption) to purification and quantification of DNA.
Any kind of help will be appreciated.
Best Regards
Arush
I can provide you a simple protocol for fungal DNA extraction.
Take 2-3 colonies directly from your culture plate using a pipette tip and dip it in a tube containing 100 microliters of nuclease free water. Vortex for a few seconds and heat in a water bath or incubator at 96 degree C for 10-15 min. Centrifuge at 13000 RPM and use 1 or 2 microliters of the supernatant for PCR.
Hi Arush,
I work with the budding yeast mostly and I've done several methods before. My preference is for the phenol-chloroform method with quantification in a nanodrop spectrophotometer.
I can provide you a simple protocol for fungal DNA extraction.
Take 2-3 colonies directly from your culture plate using a pipette tip and dip it in a tube containing 100 microliters of nuclease free water. Vortex for a few seconds and heat in a water bath or incubator at 96 degree C for 10-15 min. Centrifuge at 13000 RPM and use 1 or 2 microliters of the supernatant for PCR.
Grinding in liquid nitrogen is good so also is using Beads. Depending on the availability of materials, equipment and reagents at your disposal. Isoamylphenolchloroform extraction at ratios 25:24:1 also is works fine.
You can download these journals to help you further digest the protocols.
1) DOI: https://doi.org/10.1007/978-1-4939-7804-5_2
2) https://doi.org/10.1111/j.1472-765X.2010.02867.x
3)10.5423/PPJ.2009.25.1.108
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