QC Report


general
Report generated at2021-03-30 01:15:17
Titlegabpa_mk562
Descriptionchipseq of gabpa_mk562
Pipeline versionv1.3.6
Pipeline typetf
Genomehg38
Alignerbowtie2
Sequencing endedness{'rep1': {'paired_end': False}, 'rep2': {'paired_end': False}, 'rep3': {'paired_end': False}, 'rep4': {'paired_end': False}, 'ctl1': {'paired_end': False}, 'ctl2': {'paired_end': False}}
Peak callermacs2

Alignment quality metrics


SAMstat (raw unfiltered BAM)

rep1rep2rep3rep4ctl1ctl2
Total Reads150751851187911314949754211233563416076620407254
Total Reads (QC-failed)000000
Duplicate Reads000000
Duplicate Reads (QC-failed)000000
Mapped Reads128198991015919912838496178560553326846419919315
Mapped Reads (QC-failed)000000
% Mapped Reads85.085.585.984.597.3999999999999997.6
Paired Reads000000
Paired Reads (QC-failed)000000
Read1000000
Read1 (QC-failed)000000
Read2000000
Read2 (QC-failed)000000
Properly Paired Reads000000
Properly Paired Reads (QC-failed)000000
% Properly Paired Reads0.00.00.00.00.00.0
With itself000000
With itself (QC-failed)000000
Singletons000000
Singletons (QC-failed)000000
% Singleton0.00.00.00.00.00.0
Diff. Chroms000000
Diff. Chroms (QC-failed)000000

Marking duplicates (filtered BAM)

rep1rep2rep3rep4ctl1ctl2
Unpaired Reads11268894894930511291400156627142822904616923129
Paired Reads000000
Unmapped Reads000000
Unpaired Duplicate Reads11354137876681006091165798624489371362789
Paired Duplicate Reads000000
Paired Optical Duplicate Reads000000
% Duplicate Reads10.07568.80148.910210.5856000000000018.67528.0528

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3rep4ctl1ctl2
Total Reads11268894894930511291400156627142822904616923129
Total Reads (QC-failed)000000
Duplicate Reads000000
Duplicate Reads (QC-failed)000000
Mapped Reads11268894894930511291400156627142822904616923129
Mapped Reads (QC-failed)000000
% Mapped Reads100.0100.0100.0100.0100.0100.0
Paired Reads000000
Paired Reads (QC-failed)000000
Read1000000
Read1 (QC-failed)000000
Read2000000
Read2 (QC-failed)000000
Properly Paired Reads000000
Properly Paired Reads (QC-failed)000000
% Properly Paired Reads0.00.00.00.00.00.0
With itself000000
With itself (QC-failed)000000
Singletons000000
Singletons (QC-failed)000000
% Singleton0.00.00.00.00.00.0
Diff. Chroms000000
Diff. Chroms (QC-failed)000000

Filtered and duplicates removed


Sequence quality metrics (filtered/deduped BAM)

rep1
rep1
rep2
rep2
rep3
rep3
rep4
rep4

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)

rep1rep2rep3rep4ctl1ctl2
Total Fragments10986820873338711000413152445282605002715632616
Distinct Fragments10111032813935310263303139827382575664215536775
Positions with Two Read42095227968634621858954825006974630
NRF = Distinct/Total0.9202870.9319810.9329930.917230.9887380.993869
PBC1 = OneRead/Distinct0.9432740.9521310.9530740.9422310.9900550.99506
PBC2 = OneRead/TwoRead22.65690427.70867328.25297922.347575101.973839207.155608

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
rep1_vs_rep4
rep1_vs_rep4
rep2_vs_rep3
rep2_vs_rep3
rep2_vs_rep4
rep2_vs_rep4
rep3_vs_rep4
rep3_vs_rep4
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
rep4-pr1_vs_rep4-pr2
rep4-pr1_vs_rep4-pr2
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2

Reproducibility QC and peak detection statistics

overlapidr
Nt172008019
N1132275978
N2127376084
N3118875806
N4135512526
Np217065116
N optimal217068019
N conservative172008019
Optimal Setpooled-pr1_vs_pooled-pr2rep1_vs_rep4
Conservative Setrep1_vs_rep4rep1_vs_rep4
Rescue Ratio1.26197674418604661.5674354964816262
Self Consistency Ratio1.13998485740725152.4085510688836105
Reproducibility Testpassborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3rep4
Number of peaks29683332922576227043

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

Peak calling statistics


Peak region size

rep1rep2rep3rep4idr_optoverlap_opt
Min size210.0215.0260.0255.0237.0235.0
25 percentile233.0215.0291.0298.0682.0433.0
50 percentile (median)329.0291.0407.0420.0940.0577.0
75 percentile521.0474.0624.0626.01347.0859.0
Max size4909.04582.04614.04573.04873.04873.0
Mean449.2894586126739417.3429352396972527.1118701964133532.35203194911811052.2219728145653719.7464756288584

rep1
rep1
rep2
rep2
rep3
rep3
rep4
rep4
idr_opt
idr_opt
overlap_opt
overlap_opt

Enrichment / Signal-to-noise ratio


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

rep1rep2rep3rep4
Number of Subsampled Reads1098682087333871100041315000000
Estimated Fragment Length210215260255
Cross-correlation at Estimated Fragment Length0.3113742461369330.3071921629848640.3124873283402060.321756914810112
Phantom Peak55555555
Cross-correlation at Phantom Peak0.22811940.21611160.22024690.2407842
Argmin of Cross-correlation1500150015001500
Minimum of Cross-correlation0.098534330.081478260.10037480.1291353
NSC (Normalized Strand Cross-correlation coeff.)3.1600583.7702343.1132062.491626
RSC (Relative Strand Cross-correlation coeff.)1.6424721.6765081.769491.725244


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
rep4
rep4

Jensen-Shannon distance (filtered/deduped BAM)

rep1rep2rep3rep4
AUC0.142623901461480740.129912147747984350.146998651064627670.16272972025202972
Synthetic AUC0.48514721485957750.48333921850626840.485162432101089060.4874118172204721
X-intercept0.368523753733700670.43433085286320360.359654840115639750.27658574815964043
Synthetic X-intercept5.431627859921942e-384.7645129304390755e-304.515915389430275e-383.256672764389922e-53
Elbow Point0.68407994018081710.73722144693646610.67736427723152350.7226825064877422
Synthetic Elbow Point0.51453786116383630.49353693472454370.50670266869956420.49888640971542897
JS Distance0.278666555128486260.29115842578103220.271361411020995040.26142242560270856
Synthetic JS Distance0.43500230632929840.43169124218075520.42879469068025170.4348209789360988
% Genome Enriched31.5894068864035325.8811933447160432.48249609136020427.11836280974221
Diff. Enrichment27.00310943538995530.3382089705935526.49741193407150822.583472823541516
CHANCE Divergence0.25277058414092820.273899238439507250.24881079621352420.20098939583964912

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for macs2 raw peaks

rep1rep2rep3rep4rep1-pr1rep2-pr1rep3-pr1rep4-pr1rep1-pr2rep2-pr2rep3-pr2rep4-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.234983131441293180.247718454114593260.226313388950882960.2159650619937260.228120878588440.254944238134219570.219789397240377630.211185239033286310.22788181342374860.255328682543357550.219501567564695240.21096190609111550.243635519844023760.23636826465625580.23619825969098143

FRiP for overlap peaks

rep1_vs_rep2rep1_vs_rep3rep1_vs_rep4rep2_vs_rep3rep2_vs_rep4rep3_vs_rep4rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2rep4-pr1_vs_rep4-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.223094869229753470.223045793832496620.224776745630429430.222031279237886850.223266092548822010.223336939191427830.215912936975003940.226017439343055130.208663938926970970.19985431643583610.23213180155062568

FRiP for IDR peaks

rep1_vs_rep2rep1_vs_rep3rep1_vs_rep4rep2_vs_rep3rep2_vs_rep4rep3_vs_rep4rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2rep4-pr1_vs_rep4-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.198509706318619560.198522510439545330.200192133890063850.197050524107223650.19913748982374470.199605221817297780.190871881481891660.203105157327859530.185941778698832740.141179747009362480.18496173804324584

For macs2 raw peaks:


For overlap/IDR peaks: