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What is the typical host-to-bug DNA ratio found in nanopore sequencing without amplification?

Bioinformatics Asked by TimD1 on March 24, 2021

I’m interested in sequencing a sputum sample using an ONT MinION without performing any type of DNA amplification.
Has anyone found a good reference (or anecdotal evidence) for a range of human to pathogen DNA ratios typically found in sputum samples?
Does this depend on whether the patient is infected by a virus or bacteria, and the bacterial species?
Can I expect to find 10x as much human DNA as pathogen DNA, or 100x-1000x? I would really just like a ballpark estimate.

One Answer

My recollection of a conversation I had with Justin O'Grady is that it's about 99% host DNA for unfiltered samples. There are a few ways to reduce that portion prior to sequencing; he has written a paper that mentions a saponin-based host depletion method that is quite effective:

https://doi.org/10.1038/s41587-019-0156-5

Answered by gringer on March 24, 2021

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