QC Report


general
Report generated at2022-12-19 22:26:35
Titlemep-1_OP102_L4larva_1_1
DescriptionENCSR265QDO
Pipeline versionv1.3.6
Pipeline typetf
GenomeWS245chr
Alignerbwa
Sequencing endedness{'rep1': {'paired_end': False}, 'rep2': {'paired_end': False}, 'rep3': {'paired_end': False}, 'ctl1': {'paired_end': False}}
Peak callerspp

Alignment quality metrics


Marking duplicates (filtered BAM)

rep1rep2rep3ctl1
Unpaired Reads3445470404120013643136980187
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads729325854652172770496953
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads21.167621.14850000000000212.6634999999999997.1194999999999995

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads2716145318654811915436483234
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads2716145318654811915436483234
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 Fragments3351613393200513401896809810
Distinct Fragments2694229316368511790376459250
Positions with Two Read435690504900124688301612
NRF = Distinct/Total0.803860.8045980.8797540.948521
PBC1 = OneRead/Distinct0.8022850.8042110.8800110.950279
PBC2 = OneRead/TwoRead4.9611885.0391568.32129820.350954

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
Nt257495606
N1183583539
N2201553501
N3106191933
Np275336510
N optimal275336510
N conservative257495606
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep2rep1_vs_rep2
Rescue Ratio1.06928424404831261.1612557973599715
Self Consistency Ratio1.8980129955739711.8308329022245216
Reproducibility Testpasspass

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks395604295829599

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size115.0119.0111.0115.0115.0
25 percentile460.0476.0444.0353.0460.0
50 percentile (median)460.0476.0444.0460.0460.0
75 percentile460.0476.0444.0460.0460.0
Max size1298.01347.0765.013297.013297.0
Mean454.67692113245704471.70478141440475440.28281360856784457.83533026113673459.07779755202847

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 Reads335161339320051340189
Estimated Fragment Length175175175
Cross-correlation at Estimated Fragment Length0.5271970781449130.5561165605566950.355765269410071
Phantom Peak404040
Cross-correlation at Phantom Peak0.50336040.53774830.334238
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.46157210.50217410.2981634
NSC (Normalized Strand Cross-correlation coeff.)1.1421771.1074181.193189
RSC (Relative Strand Cross-correlation coeff.)1.5704151.5163361.596744


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
AUC0.247108925830710290.26873815696959090.21885365118214736
Synthetic AUC0.493206147236920130.493728624090082950.4897345001503545
X-intercept0.049539662459044520.044768673900637330.09646136716458414
Synthetic X-intercept1.9553127716174093e-1857.1262433716555e-2181.6398375985467922e-80
Elbow Point0.70964414132182920.69701528871364890.7014251108441717
Synthetic Elbow Point0.50556453797609250.50776465790357880.4915156256548443
JS Distance0.251208831361240340.218242026177934530.2840662795634995
Synthetic JS Distance0.35651463224063880.32609325497472850.3765172862657871
% Genome Enriched24.89893326203123225.26867988682121824.410061577878256
Diff. Enrichment30.5969470933233226.9294722703360834.82361033172099
CHANCE Divergence0.26305761036807770.231791396311616880.29941245645630304

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for spp raw peaks

rep1rep2rep3rep1-pr1rep2-pr1rep3-pr1rep1-pr2rep2-pr2rep3-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.42827500004602110.41599247838099410.425486113384074270.41610723429447460.403154761830043060.38069261395298870.416289416172338440.40443451659915370.383657479132082650.42141902242891270.42291279204334560.4226736811895407

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 Peaks0.32950623576661390.291084480414804340.29201650466660540.29599597959608190.28340919389885230.26504624675735580.3402497464138492

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 Peaks0.161946825563739350.143880327634998330.14061415492802890.123150641810359910.112315897956032670.103085662875783760.17699129264941285

For spp raw peaks:


For overlap/IDR peaks: