Hello @Anahid_Amani_Moghada, can you please rerun your command with the --verbose flag and post both the command you ran and the entire output here? Thank you.
and here is the error:
Traceback (most recent call last):
File "/opt/conda/envs/qiime2-2021.2/lib/python3.6/site-packages/q2cli/commands.py", line 329, in call
results = action(**arguments)
File "", line 2, in emp_paired
File "/opt/conda/envs/qiime2-2021.2/lib/python3.6/site-packages/qiime2/sdk/action.py", line 245, in bound_callable
output_types, provenance)
File "/opt/conda/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 "/opt/conda/envs/qiime2-2021.2/lib/python3.6/site-packages/q2_demux/_demux.py", line 399, in emp_paired
for i, record in enumerate(seqs, start=1):
File "/opt/conda/envs/qiime2-2021.2/lib/python3.6/site-packages/q2_demux/_demux.py", line 238, in iter
_trim_description(reverse_header.description)))
ValueError: Mismatched sequence descriptions: 1 length=12, 1 length=151, and 1 length=151
Plugin error from demux:
Mismatched sequence descriptions: 1 length=12, 1 length=151, and 1 length=151
Is your data in EMP Protocol Format? If you aren't sure, can you run the head command on one of your fastq files and on your barcodes file and post the results here. Aalternatively, if you aren't sure how to do that, can you DM me one of your fastq files and your barcodes file. If I can look at your fastq headers I can tell you whether they follow EMP Protocol or not.
I looked through your output and also looked further into the code where this error is raised and the code is doing the following to your forward reads, your reverse reads, and your barcodes:
Split the header SRR2830253.1 1 length=151 on the first space so we get
id = "SRR2830253.1"
description = "1 length=151"
Then it is trying to assert that the descriptions for forward, reverse, and barcode are identical, but because the length for the barcodes is 12 they are not identical. This error will probably be resolved by removing the "length=x" from your headers.
Additionally, it looks like there is a space between the @ and the id on your header line. This doesn't appear to be causing any issues, but it does seem a bit strange, so if you see any further errors you may want to get rid of those spaces.
Another thing, it seems a bit unusual that the 3rd line of your record is " +SRR2830253.2 2 length=151" as opposed to just +. You may also have to change that, but I'm not sure. I've never seen it be anything but just + before, but I'm not sure if that's illegal or anything.