hichipper QC report

2017-12-29

hichipper, version 0.7.1.

Summary Tables

GM12878_Rep2 GM12878_Rep1
Total PETs 299478736 332279257
% Reads in Anchors 38.55 38.59
% HQ Unique Mapped 87.67 86.26
% Long Range Interaction 15.63 14.46
% in Loops 0.93 0.88

The first summary table notes how many PETs are in each sample and what fraction were mapped with high-quality. First, the “% Reads in Anchors” takes all unique, mappable pairs (whether they are valid, dangling, self-circle, etc. doesn’t matter) and determines what fraction have both overlapping with anchor loci. Note, this numer should give an indication of the ChIP quality, but it may be sensitive to how peaks are determined with padding, etc. While the “% HQ Unique Mapped” reads should be self explanatory, the “% Long Range Interaction” value represents the number of intrachromosomal reads longer than the minimum length (default 5Kb) specified by the user and is a good approximation for the efficacy of the in situ ligation. Finally, the “% in Loops” is the fraction of total reads that are in loops, defined by high-quality, mapped intrachromosomal, anchor-mapped, between min and max lengths (default 5Kb and 2Mb). If the proportion of loops reads is low but the proportion of long range interactions is high, this suggests that the ChIP efficiency is relatively low.


GM12878_Rep2 GM12878_Rep1
Intrachromasomal PETs 93725004 98170528
% Reads < 5Kb 39.98 41.14
% Reads in 5KB-2Mb 49.95 48.96
% Reads > 2Mb 10.07 9.90

The second summary table shows the total number of intrachromosomal pets and the distance that they span binned by the max and min user defined distances. Only reads in the middle are used when making loops but all are used for anchor calling. If the very long interactions are disproportionately abundant, the proximity ligation step may not have been effective. In contrast, if the short range interactions are disproportionately abundant, issues with the in situ ligation likely occurred in the experiment.


Loop Width Distribution Histogram

Above is a histogram of the loop widths greater than the minimum distance supplied in the parameters. Note that the loops smaller than the minimum distance have been filtered to make the histogram more interpretable (refer to the table above to see the proportion of very loops below the minimum distance, which are likely self-ligations).

Raw Read Stats

GM12878_Rep2 GM12878_Rep1
Total_PETs 299,478,736 332,279,257
Mapped_unique_quality 262,546,720 286,612,971
Mapped_unique_quality_valid 102,943,458 107,833,087
Valid_Intra 93,725,004 98,170,528
Valid_Intra_<5Kb 37,473,831 40,389,599
Valid_Intra_5Kb-2Mb 46,810,968 48,060,076
Valid_Intra_>2Mb 9,440,205 9,720,853
Valid_Intra_anchor_5Kb-2Mb 2,783,661 2,937,509
HQ_Reads_In_Anchors 115,462,113 128,211,112

Proportional Read Stats

GM12878_Rep2 GM12878_Rep1
Total_PETs 100.00 100.00
Mapped_unique_quality 87.67 86.26
Mapped_unique_quality_valid 34.37 32.45
Valid_Intra 31.30 29.54
Valid_Intra_<5Kb 12.51 12.16
Valid_Intra_5Kb-2Mb 15.63 14.46
Valid_Intra_>2Mb 3.15 2.93
Valid_Intra_anchor_5Kb-2Mb 0.93 0.88
Long_Range_Interactions 15.63 14.46

Number of Anchor Peaks

sample npeaks
GM12878_Rep2 84598
GM12878_Rep1 84598

The number of anchor peaks used in each sample. In lowly sequenced samples, these numbers may be low and be responsible for the lack of PETs mapping into loops.