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MethPhaser

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Install:

  1. Through conda: conda install -c bioconda methphaser
  2. Through github download:
    1. Need to put the executable methphasing into ~/.bashrc for meth_phaser_parallel to call it. exact command: export "PATH=/path/to/methphaser/:$PATH"
    2. Other package requirements: check methphaser.yaml

Usage

Example

Step 1: Run MethPhaser to get block relationship. MethPhaser default ignores the largest unphased region (-ml -1), if the input data does not contain any large poorly mapped region like centromere, please use -ml -2 to not ignore any gap.

    ./meth_phaser_parallel -b test_data/HLA.R10.haplotagged.bam -r test_data/GCA_000001405.15_GRCh38_no_alt_analysis_set.chr6.fna -g test_data/LSK.filtered.gtf -vc test_data/HLA.R10.phased.vcf.gz  -o test_data/work 

Step 2: Run post processing script to get modified reads and vcf file

    ./meth_phaser_post_processing -ib test_data/HLA.R10.haplotagged.bam -if test_data/work/ -ov test_data/output.vcf -ob test_data/output -vc test_data/HLA.R10.phased.vcf.gz 
        usage: meth_phaser_parallel [-h] -b  -r  -g  -vc  [-vt] [-t] [-ml] [-c] [-a] [-o] [-k]

        methphaser: phase reads based on methlytion informaiton

        optional arguments:
          -h, --help            show this help message and exit
          -vt , --vcf_truth     Truth vcf file for benchmarking
          -t , --threads        threads
          -ml , --max_len       maximum homozygous region length for phasing, default: -1 (ignore the largest homozygous
                                region, centromere), input -2 for not skipping anything
          -c , --cut_off        the minimum percentage of vote to determine a read's haplotype
          -a , --assignment_min
                                minimum assigned read number for ranksum test
          -o , --output_dir     output_directory
          -k , --k_iterations   use at most k iterations, use -1 for unlimited iterations.

        Required arguments:
          -b , --bam_file       input methylation annotated bam file
          -r , --reference      reference genome
          -g , --gtf            gtf file from whatshap visualization
          -vc , --vcf_called    called vcf file from HapCUT2

Recomanded phasing flow:

flowchart drawio