I'm interested in utilizing the best method for large-scale gDNA extraction from composite fecal samples. Resultant gDNA will be used for Diversity profiling (16s).
I wish to take 5g of the same animal species in a mixed pool (1g each).
What gDNA extraction kits have been successful with larger sample extractions?
Also what is the consensus between using kits that use a high lysis temperature step, over a bead beating lysis step? I am looking for a protocol which is best to capture equal representation for diversity profiling these composite samples.
Any direction toward relevant literature is appreciated greatly.
Dear Christian O' Dea, high lysis temperature extraction steps have different outcomes, bead beating shears nucleic acids as well, before the large-scale extraction I would recommend to break down your testing on all the parameters you have already listed in your topical question. Consensus is very hard to find on the best method, especially if you are on your own and decided to test several strategies, but I would recommend first to use the most soft approach to open and close the eppendofs, actually, in any of the step. A Standard Operator Procedure / Protocol, as there are many, could be the consequent issue (for instance : check the literature on my research gate profile) ! I hope the sun shines on the Coast of the University of Sunshine ! Best of luck !
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