Title: Klf4

Report generated at Sat Jun 30 19:06:34 PDT 2018

Command line arguments

-species mm10 -filt_bam /users/avsec/workspace/basepair-workflow/data/Klf4/out/align/rep1/aligned_filtered.bam -peak_caller macs2 -shift_macs2 -75 -extsize_macs2 150 -nth 8 -idr_thresh 0.05 -no_dup_removal -out_dir data/Klf4 -title Klf4

Pipeline version

Latest git commit SHA1: 57cb2ff2e9cedfcf961680c36b37fe219ccf9c98 (Tue May 29 09:47:38 2018)


Directories and files
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FilesPath
Alignment
Replicate 1
Filtered & deduped bam ./out/align/rep1/aligned_filtered.bam
Tag-align ./align/rep1/aligned_filtered.tagAlign.gz
Pseudo-replicates
Replicate 1
Pseudo-replicate 1
Tag-align ./align/pseudo_reps/rep1/pr1/aligned_filtered.pr1.tagAlign.gz
Pseudo-replicate 2
Tag-align ./align/pseudo_reps/rep1/pr2/aligned_filtered.pr2.tagAlign.gz
Signal tracks
MACS2
Replicate 1
P-value ./signal/macs2/rep1/aligned_filtered.pval.signal.bw
Fold enrichment ./signal/macs2/rep1/aligned_filtered.fc.signal.bw
Peaks
MACS2
Replicate 1
Narrow peak ./peak/macs2/rep1/aligned_filtered.pval0.01.500K.narrowPeak.gz
Pseudo-replicates
Replicate 1
Pseudo-replicate 1
Narrow peak ./peak/macs2/pseudo_reps/rep1/pr1/aligned_filtered.pr1.pval0.01.500K.narrowPeak.gz
Pseudo-replicate 2
Narrow peak ./peak/macs2/pseudo_reps/rep1/pr2/aligned_filtered.pr2.pval0.01.500K.narrowPeak.gz
Naive overlap
Pseudo-replicates
Replicate 1
Overlapping peak ./peak/macs2/overlap/pseudo_reps/rep1/Klf4_rep1-pr.naive_overlap.filt.narrowPeak.gz
Optimal set
Overlapping peak ./peak/macs2/overlap/optimal_set/Klf4_rep1-pr.naive_overlap.filt.narrowPeak.gz
Conservative set
Overlapping peak ./peak/macs2/overlap/conservative_set/Klf4_rep1-pr.naive_overlap.filt.narrowPeak.gz
IDR
Pseudo-replicates
Replicate 1
IDR peak ./peak/macs2/idr/pseudo_reps/rep1/Klf4_rep1-pr.IDR0.05.filt.narrowPeak.gz
Optimal set
IDR peak ./peak/macs2/idr/optimal_set/Klf4_rep1-pr.IDR0.05.filt.narrowPeak.gz
Conservative set
IDR peak ./peak/macs2/idr/conservative_set/Klf4_rep1-pr.IDR0.05.filt.narrowPeak.gz
QC and logs
Replicate 1
Cross-corr. log ./qc/rep1/aligned_filtered.no_chrM.15M.cc.qc
Cross-corr. plot ./qc/rep1/aligned_filtered.no_chrM.15M.cc.plot.pdf
Naive overlap
Overlap QC log ./qc/Klf4_peak_overlap_final.qc
Pseudo-replicates
Replicate 1
FRiP ./peak/macs2/overlap/pseudo_reps/rep1/Klf4_rep1-pr.naive_overlap.filt.narrowPeak.FRiP.qc
IDR
IDR QC log ./qc/Klf4_IDR_final.qc
Pseudo-replicates
Replicate 1
FRiP ./peak/macs2/idr/pseudo_reps/rep1/Klf4_rep1-pr.IDR0.05.filt.narrowPeak.FRiP.qc
IDR plot ./peak/macs2/idr/pseudo_reps/rep1/Klf4_rep1-pr.unthresholded-peaks.txt.png

Visualization (add -url_base [URL_ROOT_DIR_FOR_OUT_DIR] to the command line.)
Visualize  JSON (datahub)



Workflow diagram (g./b. peak : gapped/broad peak from MACS2, n. peak : narrow peak from MACS2, r. peak : relaxed peak from SPP)


Enrichment QC (strand cross-correlation measures)
Reads15000000
Est. Fragment Len.-5
Corr. Est. Fragment Len.0.221463885794625
Phantom Peak65
Corr. Phantom Peak0.2388047
Argmin. Corr.1500
Min. Corr.0.187398
NSC1.181784
RSC0.662674

NOTE: Reads from replicates are subsampled to a max of 15M

  • Normalized strand cross-correlation coefficient (NSC) = col9 in outFile
  • Relative strand cross-correlation coefficient (RSC) = col10 in outFile
  • Estimated fragment length = col3 in outFile, take the top value


rep1
 
Number of peaks
rep1148494
rep1-pr1124843
rep1-pr2124585

  • ppr1: Raw peaks called on the first pooled pseudoreplicates
  • ppr2: Raw peaks called on the second pooled pseudoreplicates
  • repi: Raw peaks called on true replicate i
  • repi-pr1 : Raw peaks called on the first pseudoreplicate from replicate i
  • repi-pr2 : Raw peaks called on the second pseudoreplicates from replicate i
  • overlap : Overlapped peaks (filtered if blacklist exists)


Enrichment QC (Fraction of reads in raw peaks)
Fraction of Reads in Peak0.212973
Enrichment QC (Fraction of reads in overlapping peaks)
Fraction of Reads in Peak0.188135

  • ppr: Overlapping peaks comparing pooled pseudo replicates
  • rep1-pr: Overlapping peaks comparing pseudoreplicates from replicate 1
  • rep2-pr: Overlapping peaks comparing pseudoreplicates from replicate 2
  • repi-repj: Overlapping peaks comparing true replicates (rep i vs. rep j)


Reproducibility QC and Peak Detection Statistics (Overlapping peaks)
Nt0
N189465
N20
Np0
N optimal89465
N conservative89465
Optimal Setself_pseudo_rep_rep1
Conservative Setself_pseudo_rep_rep1
Rescue Ratio0.0
Self Consistency Ratio1.0
Reproducibility Testpass

  • N1: Replicate 1 self-consistent overlapping peaks (comparing two pseudoreplicates generated by subsampling Rep1 reads)
  • N2: Replicate 2 self-consistent overlapping peaks (comparing two pseudoreplicates generated by subsampling Rep2 reads)
  • Nt: True Replicate consisten overlapping peaks (comparing true replicates Rep1 vs Rep2 )
  • Np: Pooled-pseudoreplicate consistent overlapping peaks (comparing two pseudoreplicates generated by subsampling pooled reads from Rep1 and Rep2 )
  • Self-consistency Ratio: max(N1,N2) / min (N1,N2)
  • Rescue Ratio: max(Np,Nt) / min (Np,Nt)
  • Reproducibility Test: If Self-consistency Ratio >2 AND Rescue Ratio > 2, then 'Fail' else 'Pass'


Enrichment QC (Fraction of reads in IDR peaks)
Fraction of Reads in Peak0.154067

  • ppr: IDR peaks comparing pooled pseudo replicates
  • rep1-pr: IDR peaks comparing pseudoreplicates from replicate 1
  • rep2-pr: IDR peaks comparing pseudoreplicates from replicate 2
  • repi-repj: IDR peaks comparing true replicates (rep i vs. rep j)


Reproducibility QC and Peak Detection Statistics (Irreproducible Discovery Rate)
Nt0
N149174
N20
Np0
N optimal49174
N conservative49174
Optimal Setself_pseudo_rep_rep1
Conservative Setself_pseudo_rep_rep1
Rescue Ratio0.0
Self Consistency Ratio1.0
Reproducibility Testpass

  • N1: Replicate 1 self-consistent IDR 0.05 peaks (comparing two pseudoreplicates generated by subsampling Rep1 reads)
  • N2: Replicate 2 self-consistent IDR 0.05 peaks (comparing two pseudoreplicates generated by subsampling Rep2 reads)
  • Nt: True Replicate consistent IDR 0.05 peaks (comparing true replicates Rep1 vs Rep2 )
  • Np: Pooled-pseudoreplicate consistent IDR 0.05 peaks (comparing two pseudoreplicates generated by subsampling pooled reads from Rep1 and Rep2 )
  • Self-consistency Ratio: max(N1,N2) / min (N1,N2)
  • Rescue Ratio: max(Np,Nt) / min (Np,Nt)
  • Reproducibility Test: If Self-consistency Ratio >2 AND Rescue Ratio > 2, then 'Fail' else 'Pass'


rep1 pseudo-reps