Exctracting amino acid substitutions

Exctracting amino acid substitutions

0

Good day,

I’m trying to develop a pipeline to determine mutations which are responsible for amino acid changes in genes associated with antibiotic resistance. I have roughly 300 bacrtial isolates.

My approach so far has not been fruitful, in short this is what i tried:

  1. Map all my fastq sequences individually to a gene using BWA.
  2. Converted output SAM to BAM, subsequently sorted all BAM files.
  3. Run FreeBayes in order to get a vcf file with SNP’s
  4. Run VCF-annotator to get the amino acid change from the vcf file

My approach is probably flawed, but at this point I’m out of ideas. The results i get at the end are wrong.

Any help/input how I should approach this issue is greatly appriciated.


VCF


annotation


DNA


Sequencing

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