Plugin error from dada2: No reads passed the filter

Hi all,

I saw there an old post to which replies are no longer allowed, so I figured I would post my own. I’m trying to use dada2 on f and r reads, but I have some sequences that are too short. Here are my demux files.

I’m wondering if there are only a few reads that are too short, and if so - could I remove them to move forward? In the previous post, it was suggested that they use only their forward reads but I was wondering if there was another option.

Some background: My sequences are in the mixed orientation format and I’m re-processing using advice provided here. Previously I had used cutadapt to deal with the mixed orientation issue and dada2 worked fine after that. I assume the short sequences were tossed away in that process… So here is the error I’m getting now

Plugin error from dada2:

No reads passed the filter. trunc_len_f (237) or trunc_len_r (287) may be individually longer than read lengths, or trunc_len_f + trunc_len_r may be shorter than the length of the amplicon + 20 nucleotides (the length of the overlap). Alternatively, other arguments (such as max_ee or trunc_q) may be preventing reads from passing the filter.

Debug info has been saved to /var/folders/gc/t0lly6tx217bq0p81wrgmtp80000gn/T/qiime2-q2cli-err-5xu23z7q.log

What is the length of your amplicon?

They are 250 basepairs

Try truncating both fwd and rev at 150, that should be enough.
Good luck!

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