Hello I've run the following for de-multiplexing sequences:
qiime demux emp-single \
--i-seqs emp-single-end-sequences.qza \
--m-barcodes-file sample-metadata_old.tsv \
--m-barcodes-column BarcodeName \
--o-per-sample-sequences demux.qza \
--p-no-golay-error-correction \
--p-rev-comp-mapping-barcodes \
--o-error-correction-details demux-details.qza
And this is the error I got:
Plugin error from demux:
Invalid characters in sequence: ['1', '3', 'F', 'P', 'i', 'l'].
Valid characters: ['C', 'M', 'G', 'W', '-', 'H', 'T', 'K', 'S', 'Y', 'B', 'A', 'N', 'D', 'V', 'R', '.']
Note: Use `lowercase` if your sequence contains lowercase characters not in the sequence's alphabet.
Debug info has been saved to /tmp/qiime2-q2cli-err-hux310iu.log
The log report:
Traceback (most recent call last):
File "/home/kalonji/miniconda3/envs/qiime2/lib/python3.6/site-packages/q2cli/commands.py", line 329, in __call__
results = action(**arguments)
File "<decorator-gen-488>", line 2, in emp_single
File "/home/kalonji/miniconda3/envs/qiime2/lib/python3.6/site-packages/qiime2/sdk/action.py", line 245, in bound_ca$
output_types, provenance)
File "/home/kalonji/miniconda3/envs/qiime2/lib/python3.6/site-packages/qiime2/sdk/action.py", line 390, in _callabl$
output_views = self._callable(**view_args)
File "/home/kalonji/miniconda3/envs/qiime2/lib/python3.6/site-packages/q2_demux/_demux.py", line 255, in emp_single
barcodes, rev_comp_mapping_barcodes)
File "/home/kalonji/miniconda3/envs/qiime2/lib/python3.6/site-packages/q2_demux/_demux.py", line 229, in _make_barc$
barcode = str(skbio.DNA(barcode).reverse_complement())
File "/home/kalonji/miniconda3/envs/qiime2/lib/python3.6/site-packages/skbio/sequence/_grammared_sequence.py", line$
self._validate()
File "/home/kalonji/miniconda3/envs/qiime2/lib/python3.6/site-packages/skbio/sequence/_grammared_sequence.py", line$
list(self.alphabet)))
ValueError: Invalid characters in sequence: ['1', '3', 'F', 'P', 'i', 'l'].
Valid characters: ['M', 'G', '.', 'S', 'B', 'D', 'K', 'R', 'W', 'C', 'T', 'A', '-', 'N', 'H', 'V', 'Y']
Note: Use `lowercase` if your sequence contains lowercase characters not in the sequence's alphabet.
Any assistance will be highly appreciated.