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Using PCR primers from Barragan et al. (2016) to see if they are able to detect all pathogenic, intermediate, and saprophytic Leptospira. Source = High Leptospira Diversity in Animals and Humans Complicates the Search for Common Reservoirs of Human Disease in Rural Ecuador

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gregorysprenger/lepto-in-silico-pcr

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Lepto in silico PCR

Requirements

Create conda environment and install packages:

conda create -n genomedl
conda activate genomedl
mamba install ncbi-genome-download ispcr

Usage

Bash files were ran on UGE using qsub command:

  1. Download all 769 refseq files, make filenames readable, and ungzip.
qsub fetch_genomes.sh
  1. Create new folder in same directory as the from_NCBI directory.
mkdir primers
  1. Create file primers.tab with primer sequences and place into primers directory. Make sure spaces are tabs.
echo "LeptoPrimer    GAGTAACACGTGGGTAATCTTCCT        TTTACCCCACCAACTAGCTAATC" > primers.tab
  1. Find primers from genomes.
qsub find_primers.sh
  1. Concatenate forward and reverse primers by using concat_files function in python script.

  2. Run mafft on concatenated files.

qsub mafft.sh
  1. Analyze genome and save to file:
python3 analyze_lepto_genome.py
  1. Check lepto_in_silico.xlsx for finished analysis.

Notes

The analyze_lepto_genome.py script was written where speed was key. Time was taken afterwards to add comments, show some data, and fix a few errors in the analyze_lepto_genomes.ipynb.

The word 'qsub' is used to send scripts to the HPC (UGE). This can be substituted with 'bash' to run locally.

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Using PCR primers from Barragan et al. (2016) to see if they are able to detect all pathogenic, intermediate, and saprophytic Leptospira. Source = High Leptospira Diversity in Animals and Humans Complicates the Search for Common Reservoirs of Human Disease in Rural Ecuador

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