Dear all, Good day, currently I am practicing extracting total DNA using different fungi. My main purpose is to extract total DNA and send the extracted DNA for 16S Amplicon sequencing (ITS-1 and ITS-4)
this article is good for your question
https://www.ijcmas.com/6-3-2017/Swapan%20K.%20Tripathy,%20et%20al.pdf
I would recommend using Qiagen kits for Microbial DNA extraction.
https://www.qiagen.com/us/products/discovery-and-translational-research/dna-rna-purification/dna-purification/microbial-dna/dneasy-ultraclean-microbial-kit/#orderinginformation
With fungi the issue tends to be the rigidity of the cell wall making most methodologies and kits useless.
In our lab we have two protocols we use;
- CTAB DNA extraction with acid washed beads
(some reading material and plenty of protocols out there: http://www.fgsc.net/fgn44/weiland.html)
- Cycling of cold/heat; Article Fast and Reliable PCR Amplification from Aspergillus fumigat...
For sequencing purposes of ITS both should work.
this article is good for your question
https://www.ijcmas.com/6-3-2017/Swapan%20K.%20Tripathy,%20et%20al.pdf
Hello
Yes, Qiagen kits for Microbial DNA extraction it's a good suggest
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5491446/
Article Optimization of DNA extraction from human urinary samples fo...
Article Rapid and economical protocols for genomic and metagenomic D...
Good Luck
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