QC Report

	general
Report generated at	2019-11-20 11:51:59
Title	YJM789--SRB8
Description	chipseq of yeast YJM789 for tf SRB8
Pipeline version	v1.3.3
Pipeline type	tf
Genome	YJM789
Paired-end per replicate	[True, True]
Aligner	bowtie2
Peak caller	spp
Control paired-end per replicate	[True, True, True, True]

Alignment quality metrics

SAMstat (raw unfiltered BAM)

	rep1	rep2	ctl1	ctl2	ctl3	ctl4
Total Reads	6607354	6410742	5022862	7410926	11061052	8381776
Total Reads (QC-failed)	0	0	0	0	0	0
Duplicate Reads	0	0	0	0	0	0
Duplicate Reads (QC-failed)	0	0	0	0	0	0
Mapped Reads	4978505	4552968	3915768	5764661	8424118	5371312
Mapped Reads (QC-failed)	0	0	0	0	0	0
% Mapped Reads	75.3	71.0	78.0	77.8	76.2	64.1
Paired Reads	6607354	6410742	5022862	7410926	11061052	8381776
Paired Reads (QC-failed)	0	0	0	0	0	0
Read1	3303677	3205371	2511431	3705463	5530526	4190888
Read1 (QC-failed)	0	0	0	0	0	0
Read2	3303677	3205371	2511431	3705463	5530526	4190888
Read2 (QC-failed)	0	0	0	0	0	0
Properly Paired Reads	4174964	3726214	2796040	4317758	6012756	3063152
Properly Paired Reads (QC-failed)	0	0	0	0	0	0
% Properly Paired Reads	63.2	58.099999999999994	55.7	58.3	54.400000000000006	36.5
With itself	4828496	4418382	3839630	5654854	8256416	4415930
With itself (QC-failed)	0	0	0	0	0	0
Singletons	150009	134586	76138	109807	167702	955382
Singletons (QC-failed)	0	0	0	0	0	0
% Singleton	2.3	2.1	1.5	1.5	1.5	11.4
Diff. Chroms	3469	2548	1527	2704	3477	1976
Diff. Chroms (QC-failed)	0	0	0	0	0	0

Marking duplicates (filtered BAM)

	rep1	rep2	ctl1	ctl2	ctl3	ctl4
Unpaired Reads	0	0	0	0	0	0
Paired Reads	1563156	1371140	1232537	1905255	2642866	1350940
Unmapped Reads	0	0	0	0	0	0
Unpaired Duplicate Reads	0	0	0	0	0	0
Paired Duplicate Reads	41016	20272	3005	7226	12971	4196
Paired Optical Duplicate Reads	714	549	517	518	966	1248
% Duplicate Reads	2.6239	1.4785	0.24380000000000002	0.37929999999999997	0.4908	0.3106

Filtered out (samtools view -F 1804):

read unmapped (0x4)
mate unmapped (0x8, for paired-end)
not primary alignment (0x100)
read fails platform/vendor quality checks (0x200)
read is PCR or optical duplicate (0x400)

SAMstat (filtered/deduped BAM)

	rep1	rep2	ctl1	ctl2	ctl3	ctl4
Total Reads	3044280	2701736	2459064	3796058	5259790	2693488
Total Reads (QC-failed)	0	0	0	0	0	0
Duplicate Reads	0	0	0	0	0	0
Duplicate Reads (QC-failed)	0	0	0	0	0	0
Mapped Reads	3044280	2701736	2459064	3796058	5259790	2693488
Mapped Reads (QC-failed)	0	0	0	0	0	0
% Mapped Reads	100.0	100.0	100.0	100.0	100.0	100.0
Paired Reads	3044280	2701736	2459064	3796058	5259790	2693488
Paired Reads (QC-failed)	0	0	0	0	0	0
Read1	1522140	1350868	1229532	1898029	2629895	1346744
Read1 (QC-failed)	0	0	0	0	0	0
Read2	1522140	1350868	1229532	1898029	2629895	1346744
Read2 (QC-failed)	0	0	0	0	0	0
Properly Paired Reads	3044280	2701736	2459064	3796058	5259790	2693488
Properly Paired Reads (QC-failed)	0	0	0	0	0	0
% Properly Paired Reads	100.0	100.0	100.0	100.0	100.0	100.0
With itself	3044280	2701736	2459064	3796058	5259790	2693488
With itself (QC-failed)	0	0	0	0	0	0
Singletons	0	0	0	0	0	0
Singletons (QC-failed)	0	0	0	0	0	0
% Singleton	0.0	0.0	0.0	0.0	0.0	0.0
Diff. Chroms	0	0	0	0	0	0
Diff. Chroms (QC-failed)	0	0	0	0	0	0

Filtered and duplicates removed

Sequence quality metrics (filtered/deduped BAM)

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)

	rep1	rep2	ctl1	ctl2	ctl3	ctl4
Total Fragments	1557315	1366114	1191364	1836406	2555651	1314860
Distinct Fragments	1516526	1345974	1188897	1830268	2544930	1311276
Positions with Two Read	38753	19381	2445	6088	10557	3566
NRF = Distinct/Total	0.973808	0.985257	0.997929	0.996658	0.995805	0.997274
PBC1 = OneRead/Distinct	0.973786	0.985324	0.997934	0.99666	0.99582	0.997274
PBC2 = OneRead/TwoRead	38.107295	68.428925	485.251943	299.631242	240.057971	366.713685

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

0-0.5 is severe bottlenecking
0.5-0.8 is moderate bottlenecking
0.8-0.9 is mild bottlenecking
0.9-1.0 is no bottlenecking

Replication quality metrics

IDR (Irreproducible Discovery Rate) plots

Reproducibility QC and peak detection statistics

	overlap	idr
Nt	4159	628
N1	2064	457
N2	1938	450
Np	4271	458
N optimal	4271	628
N conservative	4159	628
Optimal Set	pooled-pr1_vs_pooled-pr2	rep1_vs_rep2
Conservative Set	rep1_vs_rep2	rep1_vs_rep2
Rescue Ratio	1.0269295503726859	1.3711790393013101
Self Consistency Ratio	1.065015479876161	1.0155555555555555
Reproducibility Test	pass	pass

Reproducibility QC

N1: Replicate 1 self-consistent peaks (comparing two pseudoreplicates generated by subsampling Rep1 reads)
N2: Replicate 2 self-consistent peaks (comparing two pseudoreplicates generated by subsampling Rep2 reads)
Ni: Replicate i self-consistent peaks (comparing two pseudoreplicates generated by subsampling RepX reads)
Nt: True Replicate consistent peaks (comparing true replicates Rep1 vs Rep2)
Np: Pooled-pseudoreplicate consistent peaks (comparing two pseudoreplicates generated by subsampling pooled reads from Rep1 and Rep2)
Self-consistency Ratio: max(N1,N2) / min (N1,N2)
Rescue Ratio: max(Np,Nt) / min (Np,Nt)
Reproducibility Test: If Self-consistency Ratio >2 AND Rescue Ratio > 2, then 'Fail' else 'Pass'

Number of raw peaks

	rep1	rep2
Number of peaks	8681	8843

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics

Peak region size

	rep1	rep2	idr_opt	overlap_opt
Min size	92.0	80.0	91.0	91.0
25 percentile	356.0	320.0	315.0	364.0
50 percentile (median)	356.0	320.0	358.0	364.0
75 percentile	356.0	320.0	364.0	364.0
Max size	712.0	707.0	756.0	756.0
Mean	352.9824904964866	317.46228655433674	336.20382165605093	359.2474830250527

Enrichment / Signal-to-noise ratio

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

	rep1	rep2
Number of Subsampled Reads	1848035	1710915
Estimated Fragment Length	160	135
Cross-correlation at Estimated Fragment Length	0.835532266666697	0.828796147696074
Phantom Peak	50	50
Cross-correlation at Phantom Peak	0.832114	0.8279376
Argmin of Cross-correlation	1500	1500
Minimum of Cross-correlation	0.8070374	0.799472
NSC (Normalized Strand Cross-correlation coeff.)	1.035308	1.036679
RSC (Relative Strand Cross-correlation coeff.)	1.136311	1.030161

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.

Normalized strand cross-correlation coefficient (NSC) = col9 in outFile
Relative strand cross-correlation coefficient (RSC) = col10 in outFile
Estimated fragment length = col3 in outFile, take the top value

Peak enrichment

Fraction of reads in peaks (FRiP)

FRiP for spp raw peaks

	rep1	rep2	rep1-pr1	rep2-pr1	rep1-pr2	rep2-pr2	pooled	pooled-pr1	pooled-pr2
Fraction of Reads in Peaks	0.4228349560487209	0.4114380531628553	0.3358981434033663	0.32381994391754043	0.3394720590747237	0.31931173142009434	0.34945430016206014	0.41762361956527794	0.4204429643077917

FRiP for overlap peaks

	rep1_vs_rep2	rep1-pr1_vs_rep1-pr2	rep2-pr1_vs_rep2-pr2	pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks	0.2651694669837327	0.19400679306765475	0.17458700628040638	0.2701412596136175

FRiP for IDR peaks

	rep1_vs_rep2	rep1-pr1_vs_rep1-pr2	rep2-pr1_vs_rep2-pr2	pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks	0.11889350812806647	0.10862437095142366	0.0985640343838184	0.10584359667637543

For spp raw peaks:

repX: Peak from true replicate X
repX-prY: Peak from Yth pseudoreplicates from replicate X
pooled: Peak from pooled true replicates (pool of rep1, rep2, ...)
pooled-pr1: Peak from 1st pooled pseudo replicate (pool of rep1-pr1, rep2-pr1, ...)
pooled-pr2: Peak from 2nd pooled pseudo replicate (pool of rep1-pr2, rep2-pr2, ...)

For overlap/IDR peaks:

repX_vs_repY: Comparing two peaks from true replicates X and Y
repX-pr1_vs_repX-pr2: Comparing two peaks from both pseudoreplicates from replicate X
pooled-pr1_vs_pooled-pr2: Comparing two peaks from 1st and 2nd pooled pseudo replicates