Title: YPD_WT_rep1

Report generated at Sun Sep 9 15:41:40 PDT 2018

Command line arguments

-out_dir ../tc2018/YPD_WT_rep1 -species saccer3 -fastq1_1 YPD_WT_rep1_R1_001.fastq.gz -fastq1_2 YPD_WT_rep1_R2_001.fastq.gz -nth 4

Pipeline version

Latest git commit SHA1: 895743e5d9f674f8aeca1987ad1226edd2eee27b (Thu Sep 6 15:46:23 2018)


Directories and files
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FilesPath
Raw reads
Replicate 1
Fastq 1 (194K) /users/annashch/data2018_miseq/YPD_WT_rep1_R1_001.fastq.gz
Fastq 2 (194K) /users/annashch/data2018_miseq/YPD_WT_rep1_R2_001.fastq.gz
Alignment
Replicate 1
Bam (359K) ./align/rep1/YPD_WT_rep1_R1_001.PE2SE.bam
Filtered & deduped bam (170K) ./align/rep1/YPD_WT_rep1_R1_001.PE2SE.nodup.bam
Tag-align ./align/rep1/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.tagAlign.gz
Pseudo-replicates
Replicate 1
Pseudo-replicate 1
Tag-align ./align/pseudo_reps/rep1/pr1/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.pr1.tagAlign.gz
Pseudo-replicate 2
Tag-align ./align/pseudo_reps/rep1/pr2/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.pr2.tagAlign.gz
Signal tracks
MACS2
Replicate 1
P-value ./signal/macs2/rep1/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.pf.pval.signal.bigwig
Fold enrichment ./signal/macs2/rep1/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.pf.fc.signal.bigwig
Peaks
MACS2
Replicate 1
Narrow peak (p-val thresh=0.01) ./peak/macs2/rep1/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.pf.pval0.01.300K.narrowPeak.gz
Pseudo-replicates
Replicate 1
Pseudo-replicate 1
Narrow peak ./peak/macs2/pseudo_reps/rep1/pr1/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.pr1.pf.pval0.01.300K.narrowPeak.gz
Pseudo-replicate 2
Narrow peak ./peak/macs2/pseudo_reps/rep1/pr2/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.pr2.pf.pval0.01.300K.narrowPeak.gz
Naive overlap
Pseudo-replicates
Replicate 1
Narrow peak ./peak/macs2/overlap/pseudo_reps/rep1/YPD_WT_rep1_rep1-pr.naive_overlap.narrowPeak.gz
Optimal set
Overlapping peak ./peak/macs2/overlap/optimal_set/YPD_WT_rep1_rep1-pr.naive_overlap.narrowPeak.gz
Conservative set
Overlapping peak ./peak/macs2/overlap/conservative_set/YPD_WT_rep1_rep1-pr.naive_overlap.narrowPeak.gz
QC and logs
Replicate 1
Filtered flagstat log ./qc/rep1/YPD_WT_rep1_R1_001.PE2SE.nodup.flagstat.qc
Bowtie2 map. log ./qc/rep1/YPD_WT_rep1_R1_001.PE2SE.align.log
Dedup. log ./qc/rep1/YPD_WT_rep1_R1_001.PE2SE.dup.qc
Bowtie2 map. flagstat log ./qc/rep1/YPD_WT_rep1_R1_001.PE2SE.flagstat.qc
PBC log ./qc/rep1/YPD_WT_rep1_R1_001.PE2SE.nodup.pbc.qc
Cross-corr. plot ./qc/rep1/YPD_WT_rep1_R1_001.PE2SE.nodup.tn5.no_chrM.25M.R1.cc.plot.pdf
Naive overlap
Naive overlap QC log ./qc/YPD_WT_rep1_peak_overlap_final.qc
Pseudo-replicates
Replicate 1
FRiP ./peak/macs2/overlap/pseudo_reps/rep1/YPD_WT_rep1_rep1-pr.naive_overlap.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 (raw) QC
Total389592
Total(QC-failed)0
Dupes0
Dupes(QC-failed)0
Mapped359615
Mapped(QC-failed)0
% Mapped92.31
Paired389592
Paired(QC-failed)0
Read1194796
Read1(QC-failed)0
Read2194796
Read2(QC-failed)0
Properly Paired309158
Properly Paired(QC-failed)0
% Properly Paired79.35
With itself359336
With itself(QC-failed)0
Singletons279
Singletons(QC-failed)0
% Singleton0.07
Diff. Chroms91
Diff. Chroms (QC-failed)0
Dup. QC
Unpaired Reads0
Paired Reads85479
Unmapped Reads0
Unpaired Dupes0
Paired Dupes478
Paired Opt. Dupes174
% Dupes/1000.005592
Flagstat (filtered) QC
Total170002
Total(QC-failed)0
Dupes0
Dupes(QC-failed)0
Mapped170002
Mapped(QC-failed)0
% Mapped100.00
Paired170002
Paired(QC-failed)0
Read185001
Read1(QC-failed)0
Read285001
Read2(QC-failed)0
Properly Paired170002
Properly Paired(QC-failed)0
% Properly Paired100.00
With itself170002
With itself(QC-failed)0
Singletons0
Singletons(QC-failed)0
% Singleton0.00
Diff. Chroms0
Diff. Chroms (QC-failed)0
Library Complexity QC
Total Read Pairs84586
Distinct Read Pairs84125
One Read Pair83745
Two Read Pairs339
NRF = Distinct/Total0.994550
PBC1 = OnePair/Distinct0.995483
PBC2 = OnePair/TwoPair247.035398

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)
Reads84111
Est. Fragment Len.0
Corr. Est. Fragment Len.0.274810288377056
Phantom Peak75
Corr. Phantom Peak0.2435529
Argmin. Corr.1500
Min. Corr.0.1982326
NSC1.386302
RSC1.6897

NOTE: One end of each read-pair is randomly selected and the reads are then randomly subsampled to a max of 25M

  • 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
rep12174
rep1-pr11797
rep1-pr21798

  • 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 overlapping peaks)
Fraction of Reads in Peak0.311844

  • 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
N11292
N20
Np0
N optimal1292
N conservative1292
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'