Title: YPD_rtt109_rep1

Report generated at Sun Sep 9 15:42:50 PDT 2018

Command line arguments

-out_dir ../tc2018/YPD_rtt109_rep1 -species saccer3 -fastq1_1 YPD_rtt109_rep1_R1_001.fastq.gz -fastq1_2 YPD_rtt109_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 (292K) /users/annashch/data2018_miseq/YPD_rtt109_rep1_R1_001.fastq.gz
Fastq 2 (292K) /users/annashch/data2018_miseq/YPD_rtt109_rep1_R2_001.fastq.gz
Alignment
Replicate 1
Bam (534K) ./align/rep1/YPD_rtt109_rep1_R1_001.PE2SE.bam
Filtered & deduped bam (218K) ./align/rep1/YPD_rtt109_rep1_R1_001.PE2SE.nodup.bam
Tag-align ./align/rep1/YPD_rtt109_rep1_R1_001.PE2SE.nodup.tn5.tagAlign.gz
Pseudo-replicates
Replicate 1
Pseudo-replicate 1
Tag-align ./align/pseudo_reps/rep1/pr1/YPD_rtt109_rep1_R1_001.PE2SE.nodup.tn5.pr1.tagAlign.gz
Pseudo-replicate 2
Tag-align ./align/pseudo_reps/rep1/pr2/YPD_rtt109_rep1_R1_001.PE2SE.nodup.tn5.pr2.tagAlign.gz
Signal tracks
MACS2
Replicate 1
P-value ./signal/macs2/rep1/YPD_rtt109_rep1_R1_001.PE2SE.nodup.tn5.pf.pval.signal.bigwig
Fold enrichment ./signal/macs2/rep1/YPD_rtt109_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_rtt109_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_rtt109_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_rtt109_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_rtt109_rep1_rep1-pr.naive_overlap.narrowPeak.gz
Optimal set
Overlapping peak ./peak/macs2/overlap/optimal_set/YPD_rtt109_rep1_rep1-pr.naive_overlap.narrowPeak.gz
Conservative set
Overlapping peak ./peak/macs2/overlap/conservative_set/YPD_rtt109_rep1_rep1-pr.naive_overlap.narrowPeak.gz
QC and logs
Replicate 1
Filtered flagstat log ./qc/rep1/YPD_rtt109_rep1_R1_001.PE2SE.nodup.flagstat.qc
Bowtie2 map. log ./qc/rep1/YPD_rtt109_rep1_R1_001.PE2SE.align.log
Dedup. log ./qc/rep1/YPD_rtt109_rep1_R1_001.PE2SE.dup.qc
Bowtie2 map. flagstat log ./qc/rep1/YPD_rtt109_rep1_R1_001.PE2SE.flagstat.qc
PBC log ./qc/rep1/YPD_rtt109_rep1_R1_001.PE2SE.nodup.pbc.qc
Cross-corr. plot ./qc/rep1/YPD_rtt109_rep1_R1_001.PE2SE.nodup.tn5.no_chrM.25M.R1.cc.plot.pdf
Naive overlap
Naive overlap QC log ./qc/YPD_rtt109_rep1_peak_overlap_final.qc
Pseudo-replicates
Replicate 1
FRiP ./peak/macs2/overlap/pseudo_reps/rep1/YPD_rtt109_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
Total584652
Total(QC-failed)0
Dupes0
Dupes(QC-failed)0
Mapped534163
Mapped(QC-failed)0
% Mapped91.36
Paired584652
Paired(QC-failed)0
Read1292326
Read1(QC-failed)0
Read2292326
Read2(QC-failed)0
Properly Paired455622
Properly Paired(QC-failed)0
% Properly Paired77.93
With itself533232
With itself(QC-failed)0
Singletons931
Singletons(QC-failed)0
% Singleton0.16
Diff. Chroms360
Diff. Chroms (QC-failed)0
Dup. QC
Unpaired Reads0
Paired Reads110552
Unmapped Reads0
Unpaired Dupes0
Paired Dupes1148
Paired Opt. Dupes444
% Dupes/1000.010384
Flagstat (filtered) QC
Total218808
Total(QC-failed)0
Dupes0
Dupes(QC-failed)0
Mapped218808
Mapped(QC-failed)0
% Mapped100.00
Paired218808
Paired(QC-failed)0
Read1109404
Read1(QC-failed)0
Read2109404
Read2(QC-failed)0
Properly Paired218808
Properly Paired(QC-failed)0
% Properly Paired100.00
With itself218808
With itself(QC-failed)0
Singletons0
Singletons(QC-failed)0
% Singleton0.00
Diff. Chroms0
Diff. Chroms (QC-failed)0
Library Complexity QC
Total Read Pairs107683
Distinct Read Pairs106570
One Read Pair105808
Two Read Pairs587
NRF = Distinct/Total0.989664
PBC1 = OnePair/Distinct0.992850
PBC2 = OnePair/TwoPair180.252129

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)
Reads106569
Est. Fragment Len.0
Corr. Est. Fragment Len.0.305109638586767
Phantom Peak70
Corr. Phantom Peak0.28289
Argmin. Corr.1500
Min. Corr.0.2364467
NSC1.290395
RSC1.478424

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
rep12126
rep1-pr11567
rep1-pr21544

  • 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 Peak0.261366

  • 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
N11169
N20
Np0
N optimal1169
N conservative1169
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'