QC Report


general
Report generated at2022-12-19 21:57:17
Titleunc-55_NC1639_L2larva_1_1
DescriptionENCSR258DIG
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 Reads47521232898855125927116542598
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads19806679553963741292796966
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads41.679632.957729.70999999999999716.907700000000002

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads2771456194345988514213745632
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads2771456194345988514213745632
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 Fragments46797632844414123338316001732
Distinct Fragments2754282192592587344513721629
Positions with Two Read6947814076281735401079415
NRF = Distinct/Total0.5885520.677090.708170.857509
PBC1 = OneRead/Distinct0.6112040.7119530.7382610.912407
PBC2 = OneRead/TwoRead2.4229613.3637733.71574311.598613

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
Nt133465893
N1111004828
N297024021
N358811869
Np144376271
N optimal144376271
N conservative133465893
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep2rep1_vs_rep2
Rescue Ratio1.08174734002697441.0641438995418293
Self Consistency Ratio1.88743410984526452.583199571963617
Reproducibility Testpassborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks299953034322895

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size96.098.0100.094.094.0
25 percentile384.0390.0400.0218.0376.0
50 percentile (median)384.0390.0400.0376.0376.0
75 percentile384.0390.0400.0376.0376.0
Max size3383.010132.06312.010705.010705.0
Mean376.2109684947491390.4693009919916400.72710198733347350.57327380003187364.7288910438457

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 Reads467976328444141233383
Estimated Fragment Length145145135
Cross-correlation at Estimated Fragment Length0.5019055165794470.4310287514916730.299349265787419
Phantom Peak404040
Cross-correlation at Phantom Peak0.47414230.40723970.2809193
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.43316830.36573610.2416185
NSC (Normalized Strand Cross-correlation coeff.)1.1586851.1785241.238933
RSC (Relative Strand Cross-correlation coeff.)1.6775821.5731781.468948


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
AUC0.288759186374042460.27181405450006590.23080384258784753
Synthetic AUC0.493273747457251330.491965954858127440.48808564949900946
X-intercept0.043769350076951170.0505542281228104840.10838438460018325
Synthetic X-intercept3.3340796137695087e-1893.325981581710734e-1321.653802133660937e-59
Elbow Point0.69999021891661530.69914385578291580.7041841079352532
Synthetic Elbow Point0.51127606338583620.504369823682920.5123382611941688
JS Distance0.20679227407394270.227548060822338080.271446533707158
Synthetic JS Distance0.304472443603660770.323032762342703560.35458868063785953
% Genome Enriched21.56150005888611423.50793565244816527.603814223292154
Diff. Enrichment22.3755411952008224.98641942192667631.714867023482185
CHANCE Divergence0.196221738213999380.216224094969258540.2717296739769245

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 Peaks0.32077687684740440.344433816200907740.36021903830119910.3267675907537410.329928066438208160.3053250212960180.327276348605209650.32966598712192390.30521430459745440.363575942173445740.329986939420984560.3305295518473387

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 Peaks0.23388619080127220.215411378848465280.212062662933609420.21480802870404580.21956933488177520.213530710326704650.2405209446975272

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 Peaks0.173378592396470240.15837035230177120.154777888867916880.150570674764455930.147681016167565150.118750437782864220.17868282412125447

For spp raw peaks:


For overlap/IDR peaks: