The primers are still present after cutadpt

Hello @yuanyuan12543,

From the cutadapt docs:

Each read will be searched for all given adapters, but only the best matching adapter is removed .

and

By default, at most one adapter sequence is removed from each read, even if multiple adapter sequences were provided. This can be changed by using the --times option (or its abbreviated form -n ). Cutadapt will then search for all the given adapter sequences repeatedly, either until no adapter match was found or until the specified number of rounds was reached.

This is probably the source of the issue. I would just remove each primer and adapter sequence you expect one by one to avoid confusion. Qiime also does not support the linked adapters logic.

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