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This page provides step-by-step command lines for estimating ancestry of 6 targeted sequencing samples (2 HapMap trios) in a principal component space generated using genome-wide SNP data from the Human Genome Diversity Project (HGDP).
 
This page provides step-by-step command lines for estimating ancestry of 6 targeted sequencing samples (2 HapMap trios) in a principal component space generated using genome-wide SNP data from the Human Genome Diversity Project (HGDP).
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== Download and decompress software package ==
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== Getting started ==
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Create a working directory:
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mkdir ancestry
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cd ancestry
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Download and decompress software package:
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wget http://www.sph.umich.edu/csg/chaolong/LASER/LASER-2.01.tar.gz
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tar xzvf LASER-2.01.tar.gz
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Set up to access data:
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source /home/chaolong/LASER-Tutorial/setup.txt
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What is in the setup.txt file:
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export GC=/home/mktrost/seqshop/gotcloud
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export BAM=/home/chaolong/LASER-Tutorial/BAM
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export REF=/home/chaolong/LASER-Tutorial/reference
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export HGDP=/home/chaolong/LASER-Tutorial/HGDP
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== Preparing input files for LASER ==
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Step 0: vcf --> geno
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This step prepares the reference panel by converting a VCF genotype file to a GENO file. We will skip this step and use a ready-to-use HGDP reference panel. A typical command to run the vcf2geno tool is given in the file "./LASER-2.01/vcf2geno/cmd.sh".
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Step 1: bam --> pileup
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This step uses samtools to generate pileup files from bam files.
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Please only try one sample so that we won't overload the sever with everyone running 6 jobs at the same time. Pileup files for these 6 samples have been prepared for later steps.
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It takes about 2 mins for each pileup job.
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$GC/bin/samtools mpileup -q 30 -Q 20 -f ./reference/hs37d5.fa.rz -l ./HGDP/HGDP_938.bed ./BAM/121101035.recal.bam > 121101035.recal.pileup &
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# $GC/bin/samtools mpileup -q 30 -Q 20 -f ./reference/hs37d5.fa.rz -l ./HGDP/HGDP_938.bed ./BAM/121101043.recal.bam > 121101043.recal.pileup &
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# $GC/bin/samtools mpileup -q 30 -Q 20 -f ./reference/hs37d5.fa.rz -l ./HGDP/HGDP_938.bed ./BAM/121101050.recal.bam > 121101050.recal.pileup &
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# $GC/bin/samtools mpileup -q 30 -Q 20 -f ./reference/hs37d5.fa.rz -l ./HGDP/HGDP_938.bed ./BAM/121101052.recal.bam > 121101052.recal.pileup &
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# $GC/bin/samtools mpileup -q 30 -Q 20 -f ./reference/hs37d5.fa.rz -l ./HGDP/HGDP_938.bed ./BAM/121101415.recal.bam > 121101415.recal.pileup &
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# $GC/bin/samtools mpileup -q 30 -Q 20 -f ./reference/hs37d5.fa.rz -l ./HGDP/HGDP_938.bed ./BAM/121101861.recal.bam > 121101861.recal.pileup &
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Step 2: pileup --> seq
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This step will generate a file called "hapmap_trios.seq", containing the information of 6 samples. It takes about 30 seconds to run.
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We used the pre-generated pileup files in the $BAM folder.
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python ./LASER-2.01/pileup2seq/pileup2seq.py \
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-m $HGDP/HGDP_938.site \
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-b $BAM/AMD_roi_1-based.bed \
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-i $BAM/AMD_hapmap_trios_id.txt \
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-o hapmap_trios \
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$BAM/121101035.recal.pileup \
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$BAM/121101043.recal.pileup \
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$BAM/121101050.recal.pileup \
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$BAM/121101052.recal.pileup \
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$BAM/121101415.recal.pileup \
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$BAM/121101861.recal.pileup &
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In the above command, -i specifies alternative IDs for the BAM files to be used in the .seq file (including popID and indivID). -b and -i are optional.
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