I am currently embarking on the transition from QIIME 1 to QIIME 2 and using a tutorial based off of the QIIME 2 docs to conduct the analysis. I was attempting to demultiplex, however I received the following error after running the command to demultiplex:
Plugin error from demux:
** Mismatched sequence descriptions: N:0:1, N:0:CAGTGCATATGC, and N:0:CAGTGCATATGC**
Debug info has been saved to /tmp/qiime2-q2cli-err-0rujhiem.log
the referenced log file is copied below:
Traceback (most recent call last):
File “/home/qiime2/miniconda/envs/qiime2-2018.2/lib/python3.5/site-packages/q2cli/commands.py”, line 246, in call
results = action(**arguments)
File “”, line 2, in emp_paired
File “/home/qiime2/miniconda/envs/qiime2-2018.2/lib/python3.5/site-packages/qiime2/sdk/action.py”, line 228, in bound_callable
output_types, provenance)
File “/home/qiime2/miniconda/envs/qiime2-2018.2/lib/python3.5/site-packages/qiime2/sdk/action.py”, line 363, in callable_executor
output_views = self._callable(**view_args)
File “/home/qiime2/miniconda/envs/qiime2-2018.2/lib/python3.5/site-packages/q2_demux/_demux.py”, line 331, in emp_paired
for barcode_record, forward_record, reverse_record in seqs:
File “/home/qiime2/miniconda/envs/qiime2-2018.2/lib/python3.5/site-packages/q2_demux/_demux.py”, line 211, in iter
_trim_description(reverse_header.description)))
ValueError: Mismatched sequence descriptions: N:0:1, N:0:CAGTGCATATGC, and N:0:CAGTGCATATGC
I appreciate any information/help/insight anyone has to offer regarding this error. Thank you in advanced for any guidance you have regarding this error.
This means that the record id in your barcode reads (N:0:1) don't match up or align to the record ids in your forward/reverse reads (N:0:CAGTGCATATGC), for at least one record.
How were these data processed or prepared? Did they follow the EMP protocol?
As far as I know they were prepared using EMP protocol. I downloaded the run data off of BaseSpace but previously I would download directly from the sequencer. I plan on downloading directly and running the workflow again, to see if there is any change. Do you think this will have an effect?
If the EMP protocol was used on the sequencing and data prep side of things, these IDs should all match. I would contact your sequencing center to see if they can provide a bit more detail. Keep us posted!
Sequencing was done at our lab so I was able to download the files directly from the MiSeq. After running demux using these files instead of what was downloaded from BaseSpace, I did not receive the above error or any other errors for that matter and the script ran to completion.
The only thing I can think of for the error occurring would be incorrect files downloaded from BaseSpace but I solely had the option to download the Run data and couldn’t really download much else. These files saved as Undetermined_R1 and R2 respectively, so I feel as though this is why this error occurred. @FranciscoC how did you obtain your data? Did you also download the data or was it sent from a sequencing facility ?
