omark_contextualize.py ERRORS: Max retries exceeded / Too many open files

[nam-hoang]

Dear OMArk team,

I am testing omark_contextualize.py using the provided example data and also my real data, and ran into an error which seems to have something to do with API connection Max retries exceeded/too many files opened.

16%|█████████████▋ | 412/2657 [04:17<22:35, 1.66it/s] requests.exceptions.SSLError: HTTPSConnectionPool(host='omabrowser.org', port=443): Max retries exceeded with url: /api/protein/12907256/ (Caused by SSLError(OSError(24, 'Too many open files')))

The omark_contextualize.py fragment or omark_contextualize.py missing runs with the example data (less sequences) were completed, but those with my data (more sequences) were stopped midway. The same error also occurred when I tried omark_contextualize.py assembly for both example data and my data.

I wonder if you would be able to advise me in this case? What would be the cause of this, and is there anything I could change to make this work? I would like to use this tool to improve my genome annotation.

Thank you very much and I am looking forward to hearing from you.

Best regards, Nam

[alpae]

Hi @nam-hoang ,

the error looks a lot like some temporary problems with connecting to the OMA browser API. could you provide us with some more detail how and when you run that script?

Thanks Adrian

[nam-hoang]

Thanks Adrian @alpae for your reply,

I ran the above separate commands on a Linux server (Ubuntu). Basically, the command was just like this python omark_contextualize.py fragment -m example_data/UP000005640_9606.omamer -o example_data/omark_output/ -f example_data/omark_contextualize_fragment.fa

Then, to trouble shoot, I also tested the Jupyter notebook file Contextualize_OMA.ipynb. Because I had some issue connecting my local browser to the Jupyter notebook on the server, so I ran the script within python on the server. Here, everything was smoothly until the following step where I encountered the same error 'Too many open files'. As a result, I did not get the final fasta file.

Extract uniq HOGs

uniq_HOGs = list(possible_fragments['HOG'].unique()) hog_to_medseqlen = {k: v for k, v in zip(possible_fragments['HOG'], possible_fragments['subfamily_medianseqlen'])} hog_genes = {} for hog, seq in zip(possible_fragments['HOG'],possible_fragments['gene']): glist = hog_genes.get(hog, []) glist.append(seq) hog_genes[hog] =glist hog_genes print(f'{len(hog_genes)} different HOGs')`

I later found out that it could only successfully finish for about more than 1,000 HOG sequences before getting that error, while I have a total of 2,657 uniq_HOGs. So, the way I worked around this is that I split the uniq_HOGs set into 3 subsets, and run each to write out 3 fasta files, and eventually concatenated the 3 fasta files into 1 for miniprot mapping step. For each subset, it needed to be in a fresh python terminal, or else, it would throw an error (same error as above).

Please let me know if you need any further information. Thank you very much. Nam

[alpae]

Hi @nam-hoang

indeed, it seems that the API client creates too many fresh sockets without properly cleaning them up. Fixing this requires a bit more time, but as a workaround you can simply increase the number of 'open files'. You can do this with ulimit -n in the shell before starting the python code. On linux systems, the default is usally 1024, so maybe just set it to 16000:

ulimit -n 16000
python utils/omark_contextualize.py ... 

We will try to fix this properly in the future in the omadb package.

[nam-hoang]

Thank you very much! Happy Holidays~ @alpae