Title: hd_h5

Report generated at Thu Aug 26 03:49:05 +07 2021

Command line arguments

-type histone -out_dir /mnt/scratch/ws/manuriddinov/202109231429EVL24/Vertebrates/Chicken/HD -title hd_h5 -pe -species galGal5 -system local -nth 23 -bam1 /mnt/scratch/ws/manuriddinov/202109231429EVL24/Vertebrates/Chicken/Chickseq/HD3_II_rep1.sort.bam -ctl_bam1 /mnt/scratch/ws/manuriddinov/202109231429EVL24/Vertebrates/Chicken/Chickseq/HD3_II_contrl.sort.bam

Pipeline version

Latest git commit SHA1: 0816591bacf8737379027eb3d45cd45170cfe246 (Wed Jun 6 01:27:19 2018)


Directories and files
Expand all   Collapse all
FilesPath
Alignment .
Replicate 1 .
Filtered & deduped bam (17M) ./align/rep1/HD3_II_rep1.sort.nodup.bam
Bam /mnt/scratch/ws/manuriddinov/202109231429EVL24/Vertebrates/Chicken/Chickseq/HD3_II_rep1.sort.bam
Tag-align ./align/rep1/HD3_II_rep1.sort.nodup.tagAlign.gz
Control 1 .
Filtered & deduped bam (19M) ./align/ctl1/HD3_II_contrl.sort.nodup.bam
Bam /mnt/scratch/ws/manuriddinov/202109231429EVL24/Vertebrates/Chicken/Chickseq/HD3_II_contrl.sort.bam
Tag-align ./align/ctl1/HD3_II_contrl.sort.nodup.tagAlign.gz
Pseudo-replicates .
Replicate 1 .
Pseudo-replicate 1 .
Tag-align ./align/pseudo_reps/rep1/pr1/HD3_II_rep1.sort.nodup.pr1.tagAlign.gz
Pseudo-replicate 2 .
Tag-align ./align/pseudo_reps/rep1/pr2/HD3_II_rep1.sort.nodup.pr2.tagAlign.gz
Signal tracks .
MACS2 .
Replicate 1 .
P-value ./signal/macs2/rep1/HD3_II_rep1.sort.nodup.tagAlign_x_HD3_II_contrl.sort.nodup.tagAlign.pval.signal.bw
Fold enrichment ./signal/macs2/rep1/HD3_II_rep1.sort.nodup.tagAlign_x_HD3_II_contrl.sort.nodup.tagAlign.fc.signal.bw
Peaks .
MACS2 .
Replicate 1 .
Narrow peak ./peak/macs2/rep1/HD3_II_rep1.sort.nodup.tagAlign_x_HD3_II_contrl.sort.nodup.tagAlign.pval0.01.500K.narrowPeak.gz
Pseudo-replicates .
Replicate 1 .
Pseudo-replicate 1 .
Narrow peak ./peak/macs2/pseudo_reps/rep1/pr1/HD3_II_rep1.sort.nodup.pr1.tagAlign_x_HD3_II_contrl.sort.nodup.tagAlign.pval0.01.500K.narrowPeak.gz
Pseudo-replicate 2 .
Narrow peak ./peak/macs2/pseudo_reps/rep1/pr2/HD3_II_rep1.sort.nodup.pr2.tagAlign_x_HD3_II_contrl.sort.nodup.tagAlign.pval0.01.500K.narrowPeak.gz
Naive overlap .
Pseudo-replicates .
Replicate 1 .
Overlapping peak ./peak/macs2/overlap/pseudo_reps/rep1/hd_h5_rep1-pr.naive_overlap.filt.narrowPeak.gz
Optimal set .
Overlapping peak ./peak/macs2/overlap/optimal_set/hd_h5_rep1-pr.naive_overlap.filt.narrowPeak.gz
Conservative set .
Overlapping peak ./peak/macs2/overlap/conservative_set/hd_h5_rep1-pr.naive_overlap.filt.narrowPeak.gz
QC and logs .
Fingerprint JSD Plot ./qc/hd_h5_jsd.png
Replicate 1 .
Filtered flagstat log ./qc/rep1/HD3_II_rep1.sort.nodup.flagstat.qc
JS Distance ./qc/rep1/HD3_II_rep1.sort.nodup.jsd.qc
Dedup. log ./qc/rep1/HD3_II_rep1.sort.dup.qc
PBC log ./qc/rep1/HD3_II_rep1.sort.nodup.pbc.qc
Cross-corr. log ./qc/rep1/HD3_II_rep1.sort.no_chrM.15M.R1.cc.qc
Cross-corr. plot ./qc/rep1/HD3_II_rep1.sort.no_chrM.15M.R1.cc.plot.pdf
Control 1 .
Filtered flagstat log ./qc/ctl1/HD3_II_contrl.sort.nodup.flagstat.qc
Dedup. log ./qc/ctl1/HD3_II_contrl.sort.dup.qc
PBC log ./qc/ctl1/HD3_II_contrl.sort.nodup.pbc.qc
Naive overlap .
Overlap QC log ./qc/hd_h5_peak_overlap_final.qc
Pseudo-replicates .
Replicate 1 .
FRiP ./peak/macs2/overlap/pseudo_reps/rep1/hd_h5_rep1-pr.naive_overlap.filt.narrowPeak.FRiP.qc

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)


Flagstat (filtered) QC
 rep1
ctl1
Total1704560019692948
Total(QC-failed)00
Dupes00
Dupes(QC-failed)00
Mapped1704560019692948
Mapped(QC-failed)00
% Mapped100.00100.00
Paired1704560019692948
Paired(QC-failed)00
Read185228009846474
Read1(QC-failed)00
Read285228009846474
Read2(QC-failed)00
Properly Paired1704560019692948
Properly Paired(QC-failed)00
% Properly Paired100.00100.00
With itself1704560019692948
With itself(QC-failed)00
Singletons00
Singletons(QC-failed)00
% Singleton0.000.00
Diff. Chroms00
Diff. Chroms (QC-failed)00
Dup. QC
 rep1
ctl1
Unpaired Reads00
Paired Reads3059589828058952
Unmapped Reads00
Unpaired Dupes00
Paired Dupes2207309818212478
Paired Opt. Dupes110439638220119
% Dupes/1000.721440.649079
Fingerprint JS metric
% genome enriched0.33866221252
AUC0.495971666119
CHANCE divergence0.0252020342413
Elbow Point0.0
JS Distance0.547795271969
Synthetic AUC0.50338161679
Synthetic Elbow Point0.0149788379502
Synthetic JS Distance0.233791250608
Synthetic X-intercept42.105115801
X-intercept14.9793797422
diff. enrichment0.128823910904
Fingerprint plot

Library Complexity QC
 rep1
ctl1
Total Read Pairs3053374227988047
Distinct Read Pairs86634169956892
One Read Pair19685402809024
Two Read Pairs16134982395093
NRF = Distinct/Total0.2837330.355755
PBC1 = OnePair/Distinct0.2272240.282119
PBC2 = OnePair/TwoPair1.2200451.172825

NRF (non redundant fraction)
PBC1 (PCR Bottleneck coefficient 1)
PBC2 (PCR Bottleneck coefficient 2)
PBC1 is the primary measure. Provisionally

  • 0-0.5 is severe bottlenecking
  • 0.5-0.8 is moderate bottlenecking
  • 0.8-0.9 is mild bottlenecking
  • 0.9-1.0 is no bottlenecking

Enrichment QC (strand cross-correlation measures)
Reads15000000
Est. Fragment Len.295
Corr. Est. Fragment Len.0.114590711051154
Phantom Peak150
Corr. Phantom Peak0.1194392
Argmin. Corr.1500
Min. Corr.0.1128208
NSC1.015688
RSC0.2674208

NOTE: One end of each read-pair is randomly selected and the reads are then randomly 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
rep136506
rep1-pr136552
rep1-pr236256

  • 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 Peak0.0485713
Enrichment QC (Fraction of reads in overlapping peaks)
Fraction of Reads in Peak0.00542224

  • 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
N12536
N20
Np0
N optimal2536
N conservative2536
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'