I am trying to run the "Atacama soil microbiome" tutorial on a m1x.large with paired end data that is multiplexed. I have my forward and reverse sequence files (reverse.fastq.qz, forward.fastq.qz) and my barcodes file (barcodes.fastq.qz). When I try and run the demux emp-paired command I get this error:
Plugin error from demux:
No sequences were mapped to samples. Check that your barcodes are in the correct orientation (see the rev_comp_barcodes and/or rev_comp_mapping_barcodes options).
The error log says this:
Traceback (most recent call last):
File "/home/qiime2/miniconda/envs/qiime2-2018.6/lib/python3.5/site-packages/q2cli/commands.py", line 274, in call
results = action(**arguments)
File "", line 2, in emp_paired
File "/home/qiime2/miniconda/envs/qiime2-2018.6/lib/python3.5/site-packages/qiime2/sdk/action.py", line 232, in bound_callable
File "/home/qiime2/miniconda/envs/qiime2-2018.6/lib/python3.5/site-packages/qiime2/sdk/action.py", line 367, in callable_executor
output_views = self._callable(**view_args)
File "/home/qiime2/miniconda/envs/qiime2-2018.6/lib/python3.5/site-packages/q2_demux/_demux.py", line 378, in emp_paired
raise ValueError('No sequences were mapped to samples. Check that '
ValueError: No sequences were mapped to samples. Check that your barcodes are in the correct orientation (see the rev_comp_barcodes and/or rev_comp_mapping_barcodes options).
I've run the metadata file through keemei a few times and got no errors. I've rebuilt my metadata file from scratch and tried re-running it again and I still get this error.
Going through the forums, I found users who had a similar error b/c they switched up their barcode and sequence files doing single-end analyses, but that isn't likely for me b/c I'm using paired-end analysis (Demux quits with plugin error: no Sequences mapped).
I've also demultiplexed another data-set recently and had no problem, but I did that on a different EC2 instance (m3x.large). I'm not quite sure what could be throwing this error.
I've attached my metadata file here. It's in .txt because I had to remove some patient information, but I've been uploading to QIIME2 in .tsv after running it through Keemei.
Metadata.txt (73.7 KB)