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Hi,

I really enjoyed your article! I was always hesitant to use estimators that rely on the doubleton / singleton ratio, knowing that the singleton frequency may be completely exaggerated. An earlier suggestion on how to deal with the inflation of spurious sequences comes from Robert Edgar, the author of USEARCH : he suggest to [exclude global singletons](http://www.drive5.com/usearch/manual/si...

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sequencing microbial diversity microbial ecology
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