'NoneType' object has no attribute 'strip' & Quality score length doesn't match sequence length for record

Hello! I am very new to QIIME2 and am running into an issue with my data.
This is what I have run so far:

qiime tools import \
--type 'SampleData[PairedEndSequencesWithQuality]' \
--input-format PairedEndFastqManifestPhred33V2 \
--input-path manifest.txt \
--output-path demux_seqs.qza

qiime demux summarize \
--i-data demux_seqs.qza \
--o-visualization demux_seqs.qzv

**-- output error "'NoneType' object has no attribute 'strip'"**   

I then followed along with other forums that had the same issue and proceeded with running the following to determine the line counts per sequence:

for f in *.fastq; do r=$(( $(wc -l < $f | tr -d '[:space:]') % 4 )); echo $r $f; done

Nine of my fastq files had 1 and one had 2, which I believe means that there is 1 or 2 extra lines when it’s not supposed to, likely to be blank lines (please correct me if I am wrong).
Then for each of these 10 files I ran:
grep -c ’\$’ filename.fastq

However, they all came back with 0 blank lines.

Then, I found that I could run:
qiime tools validate demux_seqs.qza
This came back with:

Result demux_seqs.qza does not appear to be valid at level=max:

 /tmp/qiime2-archive-m5btpqhe/9188183d-5361-4334-aca7-2b1bad38803c/data/41029_06_216_L001_R1_001.fastq.gz is not a(n) FastqGzFormat file:

 Quality score length doesn't match sequence length for record beginning on line 123841

I manually viewed this fastq file in bbedit and this is what I found on line 123841-123845:

@M04632:65:000000000-J8JG5:1:1111:2425:9923 1:N:0:TCTAGACT+ATATACAC

To note, these files are all unmodified from the sequencer and I am running this in qiime2-2020.6 in a computing cluster through anaconda2.
I’m at a loss for how to proceed!
Any help would be appreciated, thank you in advance!

Hi @LindsayM!

Usually when we see an error like this, it means that a file didn't completely transfer when moving from one disk to another - please double check that the files all transferred successfully. If they didn't you can reimport and try again. Let us know what you find.


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