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ClustalW using Biopython

I want to make a clustalw alignment using biopython on a fasta file that have many reads converted from fastq format to fasta format (the reads have different lengths). when I run clustalw on this file I got this error:

Traceback (most recent call last):
  File "F:\CIT656\pythonProjects\CIT656_Spring21\Project_Script.py", line 33, in <module>
    stdout, stderr = clustalw_cline()
  File "F:\CIT656\pythonProjects\CIT656_Spring21\venv\lib\site-packages\Bio\Application\__init__.py", line 574, in __call__
    raise ApplicationError(return_code, str(self), stdout_str, stderr_str)
Bio.Application.ApplicationError: Non-zero return code 1 from '"C:/Program Files (x86)/ClustalW2/Clustalw2.exe" -infile="E:\\Courses\\Bioinformatics Diploma\\Programming to bioinformatics\\Project\\CIT656-project\\Balkans\\Balkans_reads_extracts\\merged_raeds\\merged_fasta_file\\merged_reads.fasta"', message 'There was an exception in the PearsonFileParser::getSeqRange function.'
clustalw biopython fasta fastq

Could you share the code calling the clustal, and the converted fasta file? From the error message it seams that there might be an issue with the fasta file format. You can troubleshoot this be creating small fasta file (e.g. 10 sequences), and try run clustal on web, from cli, from your code to verify that it works. Then go through the actual data to see what is different there.

Btw. how big is the file?

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