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peresay/vdjbase_pipeline

By peresay

•Updated almost 4 years ago

Geno-Haplo processing pipeline used by VDJbase

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peresay/vdjbase_pipeline repository overview

⁠VDJbase Pipeline

The geno-haplo processing pipeline for VDJbase⁠.

The pipeline contain four main scripts:

⁠vdjbase-pipeline overview

The vdjbase-pipeline contains 5 sections:

  1. Initial V(D)J alignment with IMGT reference set⁠, using IgBlast⁠ and MakeDb⁠ from change-o and a custom script⁠ for collapsing similar sequences.

  2. Gene sampling and clonal inference using DefineClones⁠ from change-o. In this step it is possible to sample each gene for a representation of the repertoire. In VDJbase, for each V gene only 500 sequences (with D and J assignments) are sampled, and from these sequences, clones are inferred and one sequence per clone is taken for the downstream analysis.

  3. Novel alleles inference using TIgGER findNovelAlleles⁠ with custom gene 5' end position. For each V gene, the 5' end position bound is set to the farthest position that is present in at least 95% of the annotated sequences. The potential novel alleles are then screened for a potential chimera event. The passed alleles are added to the germline reference and the repertoire is re-annotated.

  4. Genotype inference using TIgGER inferGenotypeBayesian⁠ and creating a personal germline reference set. The repertoire is re-annotated and double chromosome deletions and haplotypes are inferred using RAbHIT deletionsByBinom and createFullHaplotype⁠.

  5. OGRDB reports using ogrdbstats⁠.

⁠Docker version

A docker for the pipeline is available. The docker is based on the immcantation container⁠ with the added VDJbase pipeline.

⁠Docker

docker pull peresay/vdjbase_pipeline

⁠Using the Container

⁠Shell inside the container

Same as in immcantation container, you can invoke a shell using the following

docker run -it peresay/vdjbase_pipeline bash
⁠Sharing files with the container

Build in the same way as the immcantation container, the same four mount points are available:

/data
/scratch
/software
/oasis
⁠Geno-Haplo pipeline: vdjbase-pipeline

Runs the vdjbase-pipeline, the are several parameters that can be modified. For the full list please go to the help guide within the docker.

# Arguments
DATA_DIR=~/P1
FASTA_FILE=/data/P1_I1_S1.fasta
SAMPLE_NAME=P1_I1_S1
NPROC=4

# Run pipeline for all avialble parameters
docker run peresay/vdjbase_pipeline -h

# Run pipeline for fasta file in docker image
docker run -v $DATA_DIR:/data:z peresay/vdjbase_pipeline \
    vdjbase-pipeline -f $FASTA_FILE -s $SAMPLE_NAME -t $NPROC

AIRR_FILE=/data/P1_I1_S1.tsv
# Run pipeline for airr format file in docker image. 
docker run -v $DATA_DIR:/data:z peresay/vdjbase_pipeline \
    vdjbase-pipeline -f $AIRR_FILE -s $SAMPLE_NAME -t $NPROC 

⁠Tools and parameters

ToolVersionparameters notes
IgBlast1.16.0up to 10 V alignments, IMGT reference set from 20/08/2020
Change-O1.0.0gene sampling up to 500 sequences, DefineClones distance threshold of 0.2,
findNovelAlleles1.0.0custom pos_range, alpha = 0.05, j_max = 0.15, min_seq = 1, germline_min = 1, min_frac = 0.75.
inferGenotypeBayesian1.0.0For all find_unmutated = FALSE; V priors 0.6, 0.4, 0.4, 0.35, 0.25, 0.25, 0.25, 0.25, 0.25; D and J - single assignment sequences with priors 0.5,0.5,0,0,0,0,0,0,0.
RAbHIT0.5.0createFullHaplotype min_minor_fraction = 0.3.
ogrdbstats0.2.0default parameters
collapse_seq1.0.0-

⁠Copying

VDJbase pipeline is free for use under the CC BY-SA 4.0

Tag summary

Content type

Image

Digest

sha256:f54e8c675…

Size

1.6 GB

Last updated

almost 4 years ago

docker pull peresay/vdjbase_pipeline