q2-cutadapt output

Good news @jessica.song! I think I solved this. :fireworks:

First, thank you for sending me the PDF describing the protocol. I noticed a key statement:

  • pairs of primers (Fw-Rev or Rev-Fw) had to be present in the sequence fragments

That is, this particular sequencing facility must expect mixed orientation reads. :rotating_light: Therefore you need to enter in both primers for each of the --p-front-* commands (in 5' - 3' orientation). :dna:
Like so:

qiime cutadapt trim-paired \
  --i-demultiplexed-sequences paired-end-demux.qza \
  --p-cores 4 \
  --p-front-f CCTACGGGNGGCWGCAG  GACTACHVGGGTATCTAAKCC \
  --p-front-r GACTACHVGGGTATCTAAKCC  CCTACGGGNGGCWGCAG \
  --p-overlap 3 \
  --p-error-rate 0.1 \
  --p-match-read-wildcards \
  --p-match-adapter-wildcards \
  --p-discard-untrimmed \
  --o-trimmed-sequences primer-trimmed-demux-2.qza \
  --verbose > cutadapt-log-2.txt

The output from the first sample looks like this:

=== Summary ===
Total read pairs processed: 133,428
Read 1 with adapter: 133,350 (99.9%)
Read 2 with adapter: 132,063 (99.0%)
Pairs written (passing filters): 131,987 (98.9%)
Total basepairs processed: 77,399,028 bp
Read 1: 38,699,572 bp
Read 2: 38,699,456 bp
Total written (filtered): 71,548,932 bp (92.4%)
Read 1: 35,774,760 bp
Read 2: 35,774,172 bp

:boom:
-Cheers!
-Mike

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