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This pipeline was developed to identify pharmacogenetic variation in publicly avalailable archaic genomes. Contact Teddy Wroblewski (tadeusz.wroblewski@cuanschutz.edu) for more information.

Required Software

  • RStudio
  • Python3
  • pypgx
  • Stargazer
  • bcftools

Genotyping pipeline

Samples were genotyped and read depth was extracted using using pypgx - version 0.1.37

pypgx bam2vcf gatk "reference_genome" "gene" "output_name.vcf" hg19 --bam_dir "${d}/bam_Files.txt"

pypgx bam2gdf hg19 "gene" "control_gene" "output_name.gdf" --bam_list "${d}/bamFiles.txt"

Examples for CYP1A2 and CYP2A6 can be found in input/genotype/ and input/depth_of_coverage/

Analysis pipeline

Genotype and read depth files were inputted into Stargazer (Lee et al. 2019) with the following command

python3 stargazer.py "genotype" -o "output_preffix" -d wgs -t "supported_gene" --vcf "input.vcf" -c "control_gene --gdf "input_rd.gdf"

Examples for CYP1A2 and CYP2A6 can be found in input/sge_output/

Stargazer

Output Processing: 2021_Archaic_PGx_SGE_Output_Analysis.R

Example output found here.

SNV Analysis

Select gene positions (start-stop gene loci + 2000bp upstream for the promoter) and filter the QUAL >= 40

bcftools view -t "chr:strt-end" "input.vcf" > "output.vcf"

bcftools view -i '%QUAL>=40' "output.vcf" > "output_filtered40.vcf"

SNVs are annotated with Annovar, CADD, SIFT, and PolyPhen2

SNV Analysis Script: 2021_Archaic_PGx_variantSNV_Analysis.R

Example output found here.

Potentially Damaging Variants

Potentially Damaging Variant Script: 2021_Archaic_PGx_potentiallyDamagingSNVs_Analysis.R

Example output found here.

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