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Error - can't makeblastdb #102

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desmodus1984 opened this issue Sep 5, 2024 · 2 comments
Open

Error - can't makeblastdb #102

desmodus1984 opened this issue Sep 5, 2024 · 2 comments

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@desmodus1984
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Hi,

I am working on an HPC with Linux. I installed MitoHifi as stated and then I loaded MitoFinder 1.4.1.

I used the following code
python MitoHiFi/src/mitohifi.py -o 5 -r CB_Parent.fasta -f c.elegans.mito.fasta -t 10 -g c.elegans.mito.gb , with PacBio Hifi reads, and I downloaded the fasta and gbk of C elegans N2, since I am working with a different strain, same species.

Everything worked fine until the makeblastdb.

2024-09-04 23:44:13 [INFO] Welcome to MitoHifi v2. Starting pipeline...
2024-09-04 23:44:13 [INFO] Length of related mitogenome is: 13794 bp
2024-09-04 23:44:13 [INFO] Number of genes on related mitogenome: 36
2024-09-04 23:44:13 [INFO] Running MitoHifi pipeline in reads mode...
2024-09-04 23:44:13 [INFO] 1. First we map your Pacbio HiFi reads to the close-related mitogenome
2024-09-04 23:44:13 [INFO] minimap2 -t 10 --secondary=no -ax map-hifi c.elegans.mito.fasta CB_Parent.fasta | samtools view -@ 10 -b -F4 -F 0x800 -o reads.HiFiMapped.bam
2024-09-04 23:44:30 [INFO] 2. Now we filter out any mapped reads that are larger than the reference mitogenome to avoid NUMTS
2024-09-04 23:44:30 [INFO] 2.1 First we convert the mapped reads from BAM to FASTA format:
2024-09-04 23:44:30 [INFO] samtools fasta reads.HiFiMapped.bam > gbk.HiFiMapped.bam.fasta
2024-09-04 23:44:31 [INFO] Total number of mapped reads: 1407
2024-09-04 23:44:31 [INFO] 2.2 Then we filter reads that are larger than 13794 bp
2024-09-04 23:44:31 [INFO] Number of filtered reads: 1348
2024-09-04 23:44:31 [INFO] 3. Now let's run hifiasm to assemble the mapped and filtered reads!
2024-09-04 23:44:31 [INFO] hifiasm --primary -t 10 -f 0 -o gbk.HiFiMapped.bam.filtered.assembled gbk.HiFiMapped.bam.filtered.fasta
2024-09-04 23:45:34 [INFO] 4. Let's run the blast of the contigs versus the close-related mitogenome
2024-09-04 23:45:34 [INFO] 4.1. Creating BLAST database:
2024-09-04 23:45:34 [INFO] makeblastdb -in c.elegans.mito.fasta -dbtype nucl
Traceback (most recent call last):
File "MitoHiFi/src/mitohifi.py", line 565, in
main()
File "MitoHiFi/src/mitohifi.py", line 207, in main
subprocess.run(makeblastdb_cmd, stderr=subprocess.DEVNULL, stdout=subprocess.DEVNULL)
File "/home/juaguila/.conda/envs/mitohifi_env/lib/python3.7/subprocess.py", line 488, in run
with Popen(*popenargs, **kwargs) as process:
File "/home/juaguila/.conda/envs/mitohifi_env/lib/python3.7/subprocess.py", line 800, in init
restore_signals, start_new_session)
File "/home/juaguila/.conda/envs/mitohifi_env/lib/python3.7/subprocess.py", line 1551, in _execute_child
raise child_exception_type(errno_num, err_msg, err_filename)
FileNotFoundError: [Errno 2] No such file or directory: 'makeblastdb': 'makeblastdb'

Any reason why this doesn't work?
I also installed Mitofinder in the same directory I am running now, and I added to the path and same problem.

@ghost
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ghost commented Sep 10, 2024

Do you have BLAST loaded as part of your script?

@desmodus1984
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Yes. I did install it in the conda environment and I also tried loading the HPC module

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