Contaminated samples or Bioinformatics problem? [16s v4 515F/806R]

Hi @Carl_Watson,

Welcome to the forum!

During your processing, did you by chance use robotic extraction or PCR? Do your blanks look more like the samples they're next to?
Well-to-well contamination is a know issue, you might want to look at this paper about it.

If you think its a cross-contamination issue (sample splashing into control), I wouldn't filter based on your negative controls. But, you may also want to check out some of the contaminant filtering threads (these were just the top hits on my search).

EDIT: One more question, what various metrics are showing no difference?

Best,
Justine

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