Unique.seqs error duplicate sequences

1

Not sure if this is a bug or not, although I have not run into this issue with analyzing previous datasets.

For some reason, I am getting the following error after running unique.seqs:

mothur > unique.seqs(fasta=current)

Using data/mothur/ptsd.trim.contigs.fasta as input file for the fasta parameter.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_14500_1657 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_15370_1735 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_16246_1756 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_17635_1764 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_11901_1874 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_16076_1920 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_16058_1929 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_16072_1939 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_13918_1940 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_18293_1940 in your fasta file, sequence names must be unique, please correct.

[ERROR]: You already have a sequence named M02779_60_000000000-K39RG_1_1101_17398_1952 in your fasta file, sequence names must be unique, please correct.

I am running Ubuntu 22.04 on an AWS EC2 instance. I did check and had similar issues with the previous few versions. This leads me to believe that perhaps I have configured something incorrectly. Thanks for your consideration.

2

Can you post the commands as you’ve entered them that were run before unique.seqs?

3

Here are the commands that were run prior to using unique.seqs:

set.seed(seed=98765)

make.contigs(file=data/references/ptsd.files, inputdir=data/raw/, outputdir=data/mothur/, maxambig=0, maxlength=275, maxhomop=8, processors=16)

4

Hmmm. Can you post the contents of ptsd.files? I wonder if there are duplicate fastq files in there

5

Thanks for helping with this issue. After inspecting my files file more closely, I noticed that I had a sample listed twice. Thanks Pat!