QC Report


general
Report generated at2020-09-18 04:10:26
TitleBZW1 (paired-end)
DescriptionBZW1
Pipeline versionv1.6.0
Pipeline typetf
Genomehg38
Alignerbowtie2
Sequencing endedness{'rep1': {'paired_end': True}, 'rep2': {'paired_end': True}, 'rep3': {'paired_end': True}, 'ctl1': {'paired_end': True}}
Peak callerspp

Alignment quality metrics


SAMstat (raw unfiltered BAM)

rep1rep2rep3ctl1
Total Reads80105608511580708538832673757676
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads76127215484206398100266072971983
Mapped Reads (QC-failed)0000
% Mapped Reads95.094.694.8999999999999998.9
Paired Reads80105608511580708538832673757676
Paired Reads (QC-failed)0000
Read140052804255790354269416336878838
Read1 (QC-failed)0000
Read240052804255790354269416336878838
Read2 (QC-failed)0000
Properly Paired Reads74653666473374787868213671457528
Properly Paired Reads (QC-failed)0000
% Properly Paired Reads93.292.592.1000000000000196.89999999999999
With itself75800424482015088054183872676994
With itself (QC-failed)0000
Singletons326791219131460822294989
Singletons (QC-failed)0000
% Singleton0.40.40.50.4
Diff. Chroms190124187990414169813052
Diff. Chroms (QC-failed)0000

Marking duplicates (filtered BAM)

rep1rep2rep3ctl1
Unpaired Reads0000
Paired Reads32802603208065583454639031673257
Unmapped Reads0000
Unpaired Duplicate Reads0000
Paired Duplicate Reads4337547250388748660165356441
Paired Optical Duplicate Reads186874122272231772227230
% Duplicate Reads13.22319999999999912.034114.08550000000000116.9116

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads56930112366053425936074852633632
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads56930112366053425936074852633632
Mapped Reads (QC-failed)0000
% Mapped Reads100.0100.0100.0100.0
Paired Reads56930112366053425936074852633632
Paired Reads (QC-failed)0000
Read128465056183026712968037426316816
Read1 (QC-failed)0000
Read228465056183026712968037426316816
Read2 (QC-failed)0000
Properly Paired Reads56930112366053425936074852633632
Properly Paired Reads (QC-failed)0000
% Properly Paired Reads100.0100.0100.0100.0
With itself56930112366053425936074852633632
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 Fragments32736011207633643447328131624735
Distinct Fragments28413454182676912962468226276588
Positions with Two Read3368828200137937152273845088
NRF = Distinct/Total0.8679570.8798040.8593520.830887
PBC1 = OneRead/Distinct0.8656470.8776160.8566810.827295
PBC2 = OneRead/TwoRead7.3010658.0104856.8310485.653576

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
Nt1953884
N11378522
N21147627
N31403934
Np25042200
N optimal25042200
N conservative1953884
Optimal Setpooled-pr1_vs_pooled-pr2pooled-pr1_vs_pooled-pr2
Conservative Setrep1_vs_rep3rep1_vs_rep3
Rescue Ratio1.28170744190807652.380952380952381
Self Consistency Ratio1.22333565702335311.5454545454545454
Reproducibility Testpassborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks298411298413298374

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size170.0164.0164.0170.0170.0
25 percentile170.0164.0164.0170.0170.0
50 percentile (median)170.0164.0164.0170.0170.0
75 percentile170.0164.0164.0170.0170.0
Max size170.0164.0164.0170.0170.0
Mean170.0164.0164.0170.0170.0

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 Reads150000001500000015000000
Estimated Fragment Length10510095
Cross-correlation at Estimated Fragment Length0.1536524468351530.1475598453423750.153657659084962
Phantom Peak505050
Cross-correlation at Phantom Peak0.15519250.14888670.155069
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.15297730.14667170.1528687
NSC (Normalized Strand Cross-correlation coeff.)1.0044131.0060561.005161
RSC (Relative Strand Cross-correlation coeff.)0.3047640.40098050.358584


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
AUC0.32189891142586760.298463659663143640.3238289631588766
Synthetic AUC0.49345323445480820.491834716567759570.49358712248955566
X-intercept0.103283644157256650.127119254022584060.10228799462589169
Synthetic X-intercept8.357599947009978e-2006.031549057200959e-1282.581348473034729e-208
Elbow Point0.51791369437957830.52092263523239820.512753510764211
Synthetic Elbow Point0.50240017705150420.50069896176613220.5089331818356684
JS Distance0.010544394512191310.0280487509610327330.01256720154184957
Synthetic JS Distance0.20429387616143470.224513348185840070.202140348089327
% Genome Enriched38.3938853523559738.7253686702280838.9908752119254
Diff. Enrichment16.24046119866011820.66103179580266416.284010539699572
CHANCE Divergence0.138850508559313550.177753861376482940.1393181680354296

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 Peaks0.06580575495793860.077707947654197570.063869882502154460.086435487778418560.104479007218181040.084130274099645780.08644651884753010.10457113634240250.084248096065096760.04896822747761910.058604855879875210.058688613582286546

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 Peaks0.0031843629444765410.00404541768800771150.0032192689783098730.00404666338966626350.0040213802673937590.0040416101225678620.005275494024370861

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 Peaks3.119109525035815e-054.147258020182869e-053.310742800530781e-051.4421190669710961e-051.9614623461242352e-052.2186378109655897e-059.517568003422348e-05

For spp raw peaks:


For overlap/IDR peaks: