QC Report


general
Report generated at2021-03-26 08:42:51
Titlemyc_k562_3
Descriptionchipseq of myc_k562_3
Pipeline versionv1.3.6
Pipeline typetf
Genomehg38
Alignerbowtie2
Sequencing endedness{'rep1': {'paired_end': False}, 'rep2': {'paired_end': False}, 'rep3': {'paired_end': False}, 'ctl1': {'paired_end': False}}
Peak callermacs2

Alignment quality metrics


SAMstat (raw unfiltered BAM)

rep1rep2rep3ctl1
Total Reads13388470122498061340725316873734
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads12403308103404271122493416202661
Mapped Reads (QC-failed)0000
% Mapped Reads92.6000000000000184.3999999999999983.796.0
Paired Reads0000
Paired Reads (QC-failed)0000
Read10000
Read1 (QC-failed)0000
Read20000
Read2 (QC-failed)0000
Properly Paired Reads0000
Properly Paired Reads (QC-failed)0000
% Properly Paired Reads0.00.00.00.0
With itself0000
With itself (QC-failed)0000
Singletons0000
Singletons (QC-failed)0000
% Singleton0.00.00.00.0
Diff. Chroms0000
Diff. Chroms (QC-failed)0000

Marking duplicates (filtered BAM)

rep1rep2rep3ctl1
Unpaired Reads98632378033187878541312964702
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads207346521841502719196330543
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads21.02219999999999827.189130.95132.5496000000000003

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads98632378033187878541312964702
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads98632378033187878541312964702
Mapped Reads (QC-failed)0000
% Mapped Reads100.0100.0100.0100.0
Paired Reads0000
Paired Reads (QC-failed)0000
Read10000
Read1 (QC-failed)0000
Read20000
Read2 (QC-failed)0000
Properly Paired Reads0000
Properly Paired Reads (QC-failed)0000
% Properly Paired Reads0.00.00.00.0
With itself0000
With itself (QC-failed)0000
Singletons0000
Singletons (QC-failed)0000
% Singleton0.00.00.00.0
Diff. Chroms0000
Diff. Chroms (QC-failed)0000

Filtered and duplicates removed


Sequence quality metrics (filtered/deduped BAM)

rep1
rep1
rep2
rep2
rep3
rep3

Open chromatin assays are known to have significant GC bias. Please take this into consideration as necessary.


Library complexity quality metrics


Library complexity (filtered non-mito BAM)

rep1rep2rep3ctl1
Total Fragments98484318006608876406512886118
Distinct Fragments77811615837646605603512614390
Positions with Two Read141420512679741444620249602
NRF = Distinct/Total0.7900910.7291040.6910080.978913
PBC1 = OneRead/Distinct0.7796340.714530.6710130.979644
PBC2 = OneRead/TwoRead4.2896613.2896352.81297449.509283

Mitochondrial reads are filtered out by default. The non-redundant fraction (NRF) is the fraction of non-redundant mapped reads in a dataset; it is the ratio between the number of positions in the genome that uniquely mapped reads map to and the total number of uniquely mappable reads. The NRF should be > 0.8. The PBC1 is the ratio of genomic locations with EXACTLY one read pair over the genomic locations with AT LEAST one read pair. PBC1 is the primary measure, and the PBC1 should be close to 1. Provisionally 0-0.5 is severe bottlenecking, 0.5-0.8 is moderate bottlenecking, 0.8-0.9 is mild bottlenecking, and 0.9-1.0 is no bottlenecking. The PBC2 is the ratio of genomic locations with EXACTLY one read pair over the genomic locations with EXACTLY two read pairs. The PBC2 should be significantly greater than 1. See more details at the ENCODE portal standard for ChIP-Seq pipeline


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


Replication quality metrics


IDR (Irreproducible Discovery Rate) plots

rep1_vs_rep2
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep3
rep2_vs_rep3
rep2_vs_rep3
rep1-pr1_vs_rep1-pr2
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep3-pr1_vs_rep3-pr2
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2

Reproducibility QC and peak detection statistics

overlapidr
Nt350166087
N11320674521
N21645603532
N31201724081
Np4758815319
N optimal4758815319
N conservative350166087
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep2_vs_rep3rep2_vs_rep3
Rescue Ratio1.35903586931688382.516674880893708
Self Consistency Ratio1.36937056885131311.2800113250283125
Reproducibility Testpassborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks499586499644499487

Top 500000 raw peaks from macs2 with p-val threshold 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size75.065.085.075.075.0
25 percentile75.065.085.0155.090.0
50 percentile (median)76.066.085.0219.0130.0
75 percentile97.081.099.0331.0201.0
Max size1461.02225.01804.02531.02531.0
Mean95.884876677889384.46107428489084105.20777918144016270.62504079900776172.1328065898966

rep1
rep1
rep2
rep2
rep3
rep3
idr_opt
idr_opt
overlap_opt
overlap_opt

Enrichment / Signal-to-noise ratio


Strand cross-correlation measures (trimmed/filtered SE BAM)

rep1rep2rep3
Number of Subsampled Reads984843180066088764065
Estimated Fragment Length756585
Cross-correlation at Estimated Fragment Length0.1081269917033540.101424357094850.098490660777091
Phantom Peak404040
Cross-correlation at Phantom Peak0.080311870.075856920.07763095
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.06878480.049219140.06352286
NSC (Normalized Strand Cross-correlation coeff.)1.5719612.0606691.550476
RSC (Relative Strand Cross-correlation coeff.)3.4130251.9598192.478564


Performed on subsampled (15000000) reads mapped from FASTQs that are trimmed to 50. Such FASTQ trimming and subsampling reads are for cross-corrleation analysis only. Untrimmed FASTQs are used for all the other analyses.

NOTE1: For SE datasets, reads from replicates are randomly subsampled to 15000000.
NOTE2: For PE datasets, the first end (R1) of each read-pair is selected and trimmed to 50 the reads are then randomly subsampled to 15000000.


rep1
rep1
rep2
rep2
rep3
rep3

Jensen-Shannon distance (filtered/deduped BAM)

rep1rep2rep3
AUC0.150419579138007460.111456270096144630.14785149628125852
Synthetic AUC0.481094593540279060.47910775315475030.47999732964362646
X-intercept0.47265907718637410.59565424279139320.4820439614048136
Synthetic X-intercept3.3463525335548415e-236.590156997027791e-191.1381032340685773e-20
Elbow Point0.65013815413282790.72168839945138220.6746200261513178
Synthetic Elbow Point0.49337473171332280.497617583935035860.489582270064137
JS Distance0.102623909765964350.119983807000323590.12130561306029806
Synthetic JS Distance0.28452701545545250.29807819362454240.2857082034168842
% Genome Enriched34.2018449874641627.42749407196810631.83123203096572
Diff. Enrichment41.7359579205230752.6042831551896743.15039654293079
CHANCE Divergence0.39125444118151640.48694103115221260.39498863365810216

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for macs2 raw peaks

rep1rep2rep3rep1-pr1rep2-pr1rep3-pr1rep1-pr2rep2-pr2rep3-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.29822815775388950.39554488150219830.307319644506183150.334272984186329060.42969764930187120.366514088009967430.33380322644616840.429444805585231070.366556969667444170.10397964727840890.132594978457257820.13234202716350008

FRiP for overlap peaks

rep1_vs_rep2rep1_vs_rep3rep2_vs_rep3rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.066068277083020930.065282986325116980.067194961126552110.116939601066059760.18315047315592180.131529502369438970.07577030771906747

FRiP for IDR peaks

rep1_vs_rep2rep1_vs_rep3rep2_vs_rep3rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.0285906101592630220.031357998326726910.032532767515220180.0217075793677065640.025990680909083780.026095415207002790.05101372892728488

For macs2 raw peaks:


For overlap/IDR peaks: