Account for mapping bias by genotype

Last updated: 2019-02-07

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File	Version	Author	Date	Message
Rmd	3fea644	Briana Mittleman	2019-02-07	add accountmapbias

We are worried there amy be false positives in the QTL analysis if the QTL is in the read and the snp leads to a mapping bias for the data. I can account for this using WASP.

I have an example script from Yang:

/project2/yangili1/yangili/TCGA_pipe/script_process.sh

STAR2.6  --genomeDir /project2/yangili1/RNAseq_pipeline/index/GRCh37/STAR_hg19 --readFilesIn $inFile\_1.fastq $inFile\_2.fastq --outSAMstrandField intronMotif --outFileNamePrefix $outFile. --outSAMtype BAM Unsorted --varVCFfile $vcfFile --waspOutputMode SAMtag --outSAMattributes vA vG

First I need to find my star indexed genome:

*/project2/gilad/briana/genome_anotation_data/star_genome

Next I need my VCF file:

/project2/gilad/briana/YRI_geno_hg19/allChrom.dose.filt.vcf.gz

runStarwWASP.sh

#!/bin/bash


#SBATCH --job-name=runStarwWASP
#SBATCH --account=pi-yangili1
#SBATCH --time=24:00:00
#SBATCH --output=runStarwWASP.out
#SBATCH --error=runStarwWASP.err
#SBATCH --partition=bigmem2
#SBATCH --mem=100G
#SBATCH --mail-type=END

module load Anaconda3
source activate three-prime-env  


in=$1 
out=$2
STAR --runThreadN 4 --genomeDir /project2/gilad/briana/genome_anotation_data/star_genome --readFilesIn $1  --outSAMstrandField intronMotif --outFileNamePrefix /project2/gilad/briana/threeprimeseq/data/STAR_bam_WASP/$2.combined.STARwWASP.bam --outSAMtype BAM Unsorted --varVCFfile /project2/gilad/briana/YRI_geno_hg19/allChrom.dose.filt.vcf   --waspOutputMode SAMtag --outSAMattributes vA vG

test_runStartwWASP.sh

#!/bin/bash


#SBATCH --job-name=test_runStarwWASP
#SBATCH --account=pi-yangili1
#SBATCH --time=24:00:00
#SBATCH --output=test_runStarwWASP.out
#SBATCH --error=test_runStarwWASP.err
#SBATCH --partition=broadwl
#SBATCH --mem=12G
#SBATCH --mail-type=END

module load Anaconda3
source activate three-prime-env  


i=/project2/gilad/briana/threeprimeseq/data/fastq/YL-SP-19239-T-combined.fastq  
describer=$(echo ${i} | sed -e 's/.*YL-SP-//' | sed -e "s/combined.fastq//")
sbatch runStarwWASP.sh $i $describer

Wraper:
wrap_runStarwWASP.sh

#!/bin/bash


#SBATCH --job-name=wrap_runStarwWASP
#SBATCH --account=pi-yangili1
#SBATCH --time=24:00:00
#SBATCH --output=wrap_runStarwWASP.out
#SBATCH --error=wrap_runStarwWASP.err
#SBATCH --partition=broadwl
#SBATCH --mem=12G
#SBATCH --mail-type=END

module load Anaconda3
source activate three-prime-env  

for i in $(ls /project2/gilad/briana/threeprimeseq/data/fastq/*);do
describer=$(echo ${i} | sed -e 's/.*YL-SP-//' | sed -e "s/combined.fastq//")
sbatch runStarwWASP.sh $i $describer
done

Quota reached at 19193N for jobs- create a wrap2
wrap_runStarwWASP2.sh

#!/bin/bash


#SBATCH --job-name=wrap_runStarwWASP2
#SBATCH --account=pi-yangili1
#SBATCH --time=24:00:00
#SBATCH --output=wrap_runStarwWASP2.out
#SBATCH --error=wrap_runStarwWASP2.err
#SBATCH --partition=broadwl
#SBATCH --mem=12G
#SBATCH --mail-type=END

module load Anaconda3
source activate three-prime-env 


for i in $(ls /project2/gilad/briana/threeprimeseq/data/fastq/YL-SP-192*); do
describer=$(echo ${i} | sed -e 's/.*YL-SP-//' | sed -e "s/combined.fastq//")
sbatch runStarwWASP.sh $i $describer
done

Sort and index these files.

SortIndexStarwWASP.sh

#!/bin/bash


#SBATCH --job-name=SortIndexStarwWASP
#SBATCH --account=pi-yangili1
#SBATCH --time=24:00:00
#SBATCH --output=SortIndexStarwWASP.out
#SBATCH --error=SortIndexStarwWASP.err
#SBATCH --partition=broadwl
#SBATCH --mem=36G
#SBATCH --mail-type=END

module load Anaconda3
source activate three-prime-env 

describer=$1

samtools sort /project2/gilad/briana/threeprimeseq/data/STAR_bam_WASP/${describer}combined.STARwWASP.bamAligned.out.bam > /project2/gilad/briana/threeprimeseq/data/STAR_bam_WASP_sort/${describer}combined.STARwWASP.bamAligned.sort.bam
samtools index /project2/gilad/briana/threeprimeseq/data/STAR_bam_WASP_sort/${describer}combined.STARwWASP.bamAligned.sort.bam

wrap_SortIndexStarwWASP.sh

#!/bin/bash


#SBATCH --job-name=wrap_SortIndexStarwWASP
#SBATCH --account=pi-yangili1
#SBATCH --time=24:00:00
#SBATCH --output=wrap_SortIndexStarwWASP.out
#SBATCH --error=wrap_SortIndexStarwWASP.err
#SBATCH --partition=broadwl
#SBATCH --mem=36G
#SBATCH --mail-type=END

module load Anaconda3
source activate three-prime-env 



for i in $(ls /project2/gilad/briana/threeprimeseq/data/STAR_bam_WASP/*STARwWASP.bamAligned.out.bam)
describer=$(echo ${i} | sed -e 's/.*YL-SP-//' | sed -e "s/combined.STARwWASP.bamAligned.out.bam//")
sbatch SortIndexStarwWASP.sh $describer
done

Session information

sessionInfo()

R version 3.5.1 (2018-07-02)
Platform: x86_64-apple-darwin15.6.0 (64-bit)
Running under: macOS  10.14.1

Matrix products: default
BLAS: /Library/Frameworks/R.framework/Versions/3.5/Resources/lib/libRblas.0.dylib
LAPACK: /Library/Frameworks/R.framework/Versions/3.5/Resources/lib/libRlapack.dylib

locale:
[1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8

attached base packages:
[1] stats     graphics  grDevices utils     datasets  methods   base     

loaded via a namespace (and not attached):
 [1] workflowr_1.1.1   Rcpp_0.12.19      digest_0.6.17    
 [4] rprojroot_1.3-2   R.methodsS3_1.7.1 backports_1.1.2  
 [7] git2r_0.23.0      magrittr_1.5      evaluate_0.11    
[10] stringi_1.2.4     whisker_0.3-2     R.oo_1.22.0      
[13] R.utils_2.7.0     rmarkdown_1.10    tools_3.5.1      
[16] stringr_1.3.1     yaml_2.2.0        compiler_3.5.1   
[19] htmltools_0.3.6   knitr_1.20

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Account for mapping bias by genotype

Briana Mittleman

2/6/2019

Session information