Hi
We are new to QIIME2-2021.2 usage… and we are not able to get correct solution for the error…
we have followed the following steps to anlyse the datas
- our input data is the following format: (Note: input fastq files are from sortmerna analysis output files – rRNA.fastq obtianed in sortmerna using the illumina WGS -demultiplexed paired end raw reads)
qiime tools import
Type: SampleData[JoinedSequencesWithQuality]
manifest file: se-33-manifest.tsv
The seqs were imported as SingleEndFastqManifestPhred33
- then quality filtered as described in the moving pictures tutorial.
- performed qiime demux summarize.
- based on the summary, decided to use –p-trim-length 150 in deblur denoise-16S step.
- submitted the qiime deblur denoise-16S with the following inputs.
Qiime deblur denoise-16S –i-demultiplexed-seqs ADS_dmux-filtered.qza –p-trim-length 150 –o-representative-sequences path/ADS_rep-seqs-deblur.qza –o-table path/ADS_table-deblur.qza –o-stats path/ADS_deblur-stats.qza –p-sample-stats –p-jobs-to-start 90 –verbose
In run, received the following error message, suggest us to solve the issue
(base) srini1@srini:~ conda activate qiime2-2021.2
(qiime2-2021.2) srini1@srini:~ qiime deblur denoise-16S --i-demultiplexed-seqs /home/srini1/prabu/Q2/QC/ADS_demux-filtered.qza --p-trim-length 150 --o-representative-sequences /home/srini1/prabu/Q2/deblur/ADS_rep-seqs-deblur.qza --o-table /home/srini1/prabu/Q2/deblur/ADS_table-deblur.qza --o-stats /home/srini1/prabu/Q2/deblur/ADS_deblur-stats.qza --p-sample-stats --p-jobs-to-start 90 --verbose
Traceback (most recent call last):
File “/home/srini1/miniconda2/envs/qiime2-2021.2/lib/python3.6/site-packages/q2cli/commands.py”, line 329, in call
results = action(**arguments)
File “”, line 2, in denoise_16S
File “/home/srini1/miniconda2/envs/qiime2-2021.2/lib/python3.6/site-packages/qiime2/sdk/action.py”, line 245, in bound_callable
output_types, provenance)
File “/home/srini1/miniconda2/envs/qiime2-2021.2/lib/python3.6/site-packages/qiime2/sdk/action.py”, line 390, in callable_executor
output_views = self._callable(**view_args)
File “/home/srini1/miniconda2/envs/qiime2-2021.2/lib/python3.6/site-packages/q2_deblur/_denoise.py”, line 100, in denoise_16S
hashed_feature_ids=hashed_feature_ids)
File “/home/srini1/miniconda2/envs/qiime2-2021.2/lib/python3.6/site-packages/q2_deblur/_denoise.py”, line 192, in _denoise_helper
“of the data being denoised.” % trim_length)
ValueError: No sequences passed the filter. It is possible the trim_length (150) may exceed the longest sequence, that all of the sequences are artifacts like PhiX or adapter, or that the positive reference used is not representative of the data being denoised.
Plugin error from deblur:
No sequences passed the filter. It is possible the trim_length (150) may exceed the longest sequence, that all of the sequences are artifacts like PhiX or adapter, or that the positive reference used is not representative of the data being denoised.
See above for debug info.
enclosed the dropbox link for the files