More barcodes were provided than sequences- output error

Technical Support
1

Hello, I receive the following error no matter which module of qiime used, here it is 2020.8, but this error is present for 2021.4 and 2022.8 as well. My metadata was validated by Keemei, it has the correct # of barcode sequences to ID's in the file, yet I keep getting this output. The column header for the barcodes is also exactly the same in the command line.

I have also tried this line "--o-per-sample-sequences demux.qza \ " instead of "--output-dir demux.qza ".
mappingfile1 2.tsv (986 Bytes)

Any help appreciated! I have not seen this error in previous forums.

code used

#!/bin/bash
#SBATCH -p normal
#SBATCH --nodes 1
#SBATCH --ntasks-per-node=24
#SBATCH --mem=700g
#SBATCH -J demux1

module purge
module load QIIME2/2020.8

qiime demux emp-single
--i-seqs emp-single-end-sequences.qza
--m-barcodes-file mappingfile.tsv
--m-barcodes-column barcodesequences
--output-dir demux.qza
--o-error-correction-details demux-details.qza
--p-no-golay-error-correction

output------------------------
When running QIIME2, do not have any other modules loaded as other modules might conflict with QIIME2. You can$
module purge
module load QIIME2/2020.8
Plugin error from demux:

More barcodes were provided than sequences.

Debug info has been saved to /tmp/qiime2-q2cli-err-nrex0po3.log

2

Hello and welcome to QIIME2 Forum,

At the moment the reason for this error is unclear, provide your input sequences, please.

Cheers,
V

3

Hello, thanks for the help~ my metadata file is stated here,
mappingfile - Sheet1-3.tsv (5.7 KB)
which includes my barcodes. Is there another file you are looking for?

thanks,
Breanna

4

Hello,

I was thinking about file emp-single-end-sequences.qza from -i--seqs parameter.

Cheers,
V

5

Hi V,

Thanks for clarifying! Here is the .qza file I have in a box link for downloading.
https://uofi.box.com/s/p7z0izpckuz9i8hxv5c3hrjv6n5bl7zh

Thanks,
Breanna

6

Hello,

It seems like there is an issue with sequencing data. How did you import it?

Cheers,
V

7

Hi V,

This is the command I used for importing. I hope this helps!

#!/bin/bash

----------------SLURM Parameters----------------

#SBATCH -p normal
#SBATCH -n 24
#SBATCH --mem=1000g
#SBATCH -N 1
#SBATCH -J import

----------------Load Modules--------------------

module load QIIME2/2020.8

----------------Commands------------------------

qiime tools import
--type EMPSingleEndSequences
--input-path emp-single-end-sequences
--output-path emp-single-end-sequences.qza

8

Hello,

Ok, I see. It looks like your sequencing data is corrupted. There are simply fewer sequences than there are barcodes.
What is the origin of data? Is it from "Moving Pictures" tutorial?

Cheers,
V

1 Like
9

Hi V,

The origin of the data is from our schools sequencing center. I have been following the Moving Pictures workflow, but am not using their tutorial data. I have tried redownloading it and nothing is different, perhaps I need to discuss with our sequencing center bc I am very stuck.

Thanks,
Breanna

10

Hello,

Data only have 20 sequences and much more barcodes. Additionally, I don't think EMP single-end protocol is that common anymore, unless you specifically planned it for the experiment.
Contact the provider for the details.

Cheers,
V

1 Like
11

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