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-   -   Metagenomic DNA/RNA from human tissue samples (http://seqanswers.com/forums/showthread.php?t=71594)

simpsoncl 09-20-2016 01:39 PM

Metagenomic DNA/RNA from human tissue samples
 
Hi,

Is anyone extracting microbial DNA/RNA from human (or animal) tissue? I've got some kidney biopsies that I want to look at the microbiome but I'm not having much luck in finding a protocol to isolate the nucleic acids. Any advice, kit recommendations etc would be gratefully received.

Thanks,

Claire

nucacidhunter 09-20-2016 02:35 PM

NEBNext microbiome DNA enrichment kit

fanli 09-20-2016 03:39 PM

Do you mean extraction or enrichment of microbial DNA/RNA? For extraction, we've been using the AllPrep kit from Qiagen with reasonable success.

@nucacidhunter, how much enrichment do you see using the NEBNext? We've had pretty limited success (~20-40%), depending on sample.

Enrichment of microbial RNA seems trickier - I haven't heard of anything besides the theoretical "you can use an array to pull out human RNA".

nucacidhunter 09-20-2016 07:35 PM

I have not used them but following paper has compared NEBNext Microbiome DNA Enrichment Kit and the Molzym MolYsis Basic kit. They use different strategies for DNA enrichment.

http://www.ncbi.nlm.nih.gov/pubmed/27237775


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