I am trying to denoise my forward and reverse reads (2x300bp).
My primers are 341f-805r, for 16s V3-V4.
Here are my codes that I've used.
!qiime dada2 denoise-paired
--p-trim-left-f 17 \
Trimming parameter 17 and 21 was determined based on length of f-primer and r-primer.
However, I got error displaying "No read pass filter". The thing is, the codes above were well-executed 2 days ago, but when I try to re-run today, it displays error.
And now, it displays the error below regardless of a truncation parameter that I choose. It says 'no reads passed the filter' when I choose 0/0, 150/150, 300/300 as truncation parameters, either.
No reads passed the filter. trunc_len_f (260) or trunc_len_r (240) may be individually longer than read lengths, or trunc_len_f + trunc_len_r may be shorter than the length of the amplicon + 12 nucleotides (the length of the overlap).
I additionally attach my quality plot. Plus, what do you think what would be a proper truncation parameter based on the quality plot below? I think forward for 280 and reverse for 220 would be great, but it would be great to hear other's opinion.