Unique.seqs ERROR after make.contigs

Hi
I am getting an error in unique.seqs command like:

[ERROR]: You already have a sequence named NB501984_42_H7FN2AFX2_2_21112_20285_13267 in your fasta file, sequence names must be unique, please correct.
[ERROR]: NB501984_42_H7FN2AFX2_2_21112_20285_13267 is not in your count table. Please correct.
12347240 801866

However after searching through my fasta file, I did not find any duplicated sequence names… and I am also not sure why the sequence that mentioned in the error was not included in my count file…?

Then I went back and tried to run summary.seqs (fasta=/Volumes/mypassport/mothur-2/EISPmothur2.trim.contigs.fasta)

Start End NBases Ambigs Polymer NumSeqs

Minimum: 1 30 30 0 2 1

2.5%-tile: 1 99 99 0 3 1019301

25%-tile: 1 141 141 0 4 10193008

Median: 1 168 168 0 4 20386015

75%-tile: 1 168 168 0 4 30579022

97.5%-tile: 1 174 174 2 6 39752729

Maximum: 1 270 270 58 135 40772029

Mean: 1 157 157 0 4

of Seqs: 40772029

It took 1202 secs to summarize 40772029 sequences.

count.groups (count=/Volumes/mypassport/mothur-2/EISPmothur2.contigs.count_table)

Total seqs: 40679445.

it seems that the sequence number does not match here?
I am using the version1.48 and 8 processors, the upstream command is only make.contigs.
Does anyone kindly know what should I do to fix this problem?

Hello
I want to add some actions I tried here…
I re-ran the make.contigs from the beggining using a 1.44 version of Mothur
and then
unique.seqs (fasta=/Volumes/mypassport/mothur-3/EISPmothur3.trim.contigs.good.fasta) (I re-ran in another directionary for the 1.44mothur)
and more errors went out as **** Exceeded maximum allowed command errors, quitting ****

[ERROR]: You already have a sequence named NB501984_42_H7FN2AFX2_1_21205_25114_6689 in your fasta file, sequence names must be unique, please correct.

so I tried the list.seqs(fasta=/Volumes/mypassport/mothur-3/EISPmothur3.trim.contigs.good.fasta)
and it ended up with tons of warnings as
[WARNING]: We found more than 25% of the bases in sequence !!!H__PAGEZEROx__TEXT?_?___text__TEXT?_Z??__stubs__TEXT to be ambiguous. Mothur is not setup to process protein sequences.

[WARNING]: We found more than 25% of the bases in sequence M?H?[A^A_]?\H?[A^A_]?f?UH??H??H??H??t to be ambiguous. Mothur is not setup to process protein sequences.

[WARNING]: We found more than 25% of the bases in sequence ?H??H??H?U?H?M?E?|$ to be ambiguous. Mothur is not setup to process protein sequences.

and my good.accnos file is zerobyte.
so, could anyone knows what was happening here…?

thanks in advance!

hi everyone
sorry to bother again!
I closely checked my stability file from the very beginning this time and found I import one of my samples twice…
I think that is where the error came from.
Thank you!!

Fanyu

Glad you found it - holler if you need any more help
Pat

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