## extract chromosome from bam file samtools view -bo test.bam /home/brandl/mnt/chip-seq_study/ChIPSeq_February_2014/alignments_trimmed_nomulti_pooled/H2Az.bam 1 ######################################################################################################################## ## macs2 playground macs2 -n --broad --gsize #macs2 -n --gsize ######################################################################################################################## ### deeptools ## https://github.com/fidelram/deepTools/wiki/All-command-line-options#bamCorrelate #bamCorrelate bins --bamfiles $bamFiles --region 10:1:100000 --plotFile="bam_correlation.png" --numberOfProcessors=4 --corMethod spearman ## see how well bam files correlate using untrimmed data bamCorrelate bins --bamfiles $bamFiles --plotFile="bam_correlation_untrimmed.png" --numberOfProcessors=4 --corMethod spearman ######################################################################################################################## ### Peak calling with SPP Rscript /home/brandl/bin/phantompeakqualtools/run_spp.R -c="../alignments_untrimmed/H2Az_Rep1_Lane2_Lib4454.0001.bam" -savp -out="test.txt" ######################################################################################################################## ### other qc ## http://www.nature.com/nmeth/journal/v11/n1/full/nmeth.2786.html # Teytelman et al. discovered that highly expressed loci were always enriched in ChIP peaks, regardless of which protein was pulled dow ######################################################################################################################### #### CHANCE mcdir $baseDir/qc_chance ## convert example to sam format bamToBed -i $baseDir/alignments_trimmed/H2Az_Rep1_Lane1_Lib4454_ca.bam > H2Az_Rep1_Lane1_Lib4454_ca.bed samtools view -h -o H2Az_Rep1_Lane1_Lib4454_ca.sam $baseDir/alignments_trimmed/H2Az_Rep1_Lane1_Lib4454_ca.bam /local/home/henry/bin/CHANCE/run_chance_linux.sh /usr/local/MATLAB/MATLAB_Compiler_Runtime/v717/ ######################################################################################################################## ## Use phantompeakqualtools to calculate quality tag (strand cross-correlation of peak) # did the antibody-treatment enrich sufficiently so that the ChIP signal can be separated from the background signal? ## --> use https://code.google.com/p/phantompeakqualtools/ mcdir $baseDir/qc_phantompeaks #Rscript /local/home/brandl/bin/phantompeakqualtools/run_spp.R #Rscript run_spp.R <bamfile> -savp -out=<outfile> ctrlBam=$baseDir/alignments_untrimmed/H3-3_Rep1_Lane1_Lib4453.bam #DEBUG ctrlBam=$baseDir/alignments_untrimmed/H3K4me3_Rep2_Lane2_Lib4455.bam for bamFile in $baseDir/alignments_untrimmed/*bam ; do echo "processing $bamFile..." # Rscript /home/brandl/bin/phantompeakqualtools/run_spp.R -c=$bamFile -savp -ou t=$(basename $bamFile).pt.log ## one output file is enough because output is tabular including inpuyt file name # Rscript /home/brandl/bin/phantompeakqualtools/run_spp.R -c=$bamFile -i=$ctrlBam -savp -savd -odir=$(pwd) -out=phantom_qc.txt ## with control # ( Rscript /home/brandl/bin/phantompeakqualtools/run_spp.R -c=$bamFile -savp -savd -odir=$(pwd) -out=phantom_qc_noctrl.txt &> $(basename $bamFile).pt.log ) & ## without control # ( Rscript /home/brandl/bin/phantompeakqualtools/run_spp.R -c=$bamFile -savp -savd -odir=$(pwd) -out=phantom_qc_noctrl.txt &> $(basename $bamFile).pt.log ) & done wait ## todo visualize results #phantom output format #format:Filename<tab>numReads<tab>estFragLen<tab>corr_estFragLen<tab>PhantomPeak<tab>corr_phantomPeak<tab>argmin_corr<tab>min_corr<tab>Normalized SCC (NSC)<tab>Relative SCC (RSC)<tab>QualityTag) mailme "phantom qc done" ######################################################################################################################## ### motifs ## run meme-chip ######################################################################################################################## ### profiles ## try deepTools ## differential binding diffbind