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        Download the raw data used to create the plots in this report below:

        Note that additional data was saved in multiqc_data when this report was generated.


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        If you use plots from MultiQC in a publication or presentation, please cite:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

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        About MultiQC

        This report was generated using MultiQC, version 1.11

        You can see a YouTube video describing how to use MultiQC reports here: https://youtu.be/qPbIlO_KWN0

        For more information about MultiQC, including other videos and extensive documentation, please visit http://multiqc.info

        You can report bugs, suggest improvements and find the source code for MultiQC on GitHub: https://github.com/ewels/MultiQC

        MultiQC is published in Bioinformatics:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

        A modular tool to aggregate results from bioinformatics analyses across many samples into a single report.

        This report has been generated by the nf-core/hicar analysis pipeline. For information about how to interpret these results, please see the documentation.

        Report generated on 2021-09-10, 19:47 based on data in: /Users/ouj/github/new/work/4e/027059f64e7bd580d64ef348043c11


        General Statistics

        Showing 24/24 rows and 6/8 columns.
        Sample NameM Reads Mapped% BP Trimmed% Dups% GCLengthM Seqs
        KD_REP1.srt
        1.2
        KD_REP1_T0_1
        54.4%
        45%
        61 bp
        0.5
        KD_REP1_T0_2
        33.3%
        50%
        71 bp
        0.5
        KD_REP1_T1_1
        32.0%
        45%
        95 bp
        0.1
        KD_REP1_T1_2
        10.2%
        48%
        90 bp
        0.1
        KD_REP2.srt
        1.1
        KD_REP2_T0_1
        54.4%
        45%
        61 bp
        0.5
        KD_REP2_T0_2
        33.3%
        50%
        71 bp
        0.5
        KD_REP2_T1_1
        22.4%
        45%
        95 bp
        0.0
        KD_REP2_T1_2
        9.5%
        48%
        90 bp
        0.0
        KD_rep1_R2
        0.0%
        KD_rep2_R2
        1.2%
        WT_REP1.srt
        1.4
        WT_REP1_T0_1
        56.4%
        45%
        61 bp
        0.6
        WT_REP1_T0_2
        36.2%
        50%
        71 bp
        0.6
        WT_REP1_T1_1
        40.6%
        46%
        95 bp
        0.1
        WT_REP1_T1_2
        12.5%
        50%
        90 bp
        0.1
        WT_REP2.srt
        1.4
        WT_REP2_T0_1
        56.4%
        45%
        61 bp
        0.6
        WT_REP2_T0_2
        36.2%
        50%
        71 bp
        0.6
        WT_REP2_T1_1
        42.4%
        46%
        95 bp
        0.1
        WT_REP2_T1_2
        12.1%
        50%
        90 bp
        0.1
        WT_rep1_R2
        1.2%
        WT_rep2_R2
        0.0%

        PAIRS QC

        Pairs QC report

        PAIRS reads stats

        Table of mapped reads stats.

        Showing 4/4 rows and 6/6 columns.
        sampletotalduplicatenon_duplicatedtranscislongRange
        KD_REP1.pairsam.stat
        581856.0
        141038.0
        225918.0
        0.0
        225918.0
        126302.0
        KD_REP2.pairsam.stat
        558774.0
        140327.0
        213162.0
        0.0
        213162.0
        119506.0
        WT_REP1.pairsam.stat
        686168.0
        178696.0
        255461.0
        0.0
        255461.0
        143013.0
        WT_REP2.pairsam.stat
        694922.0
        179219.0
        261119.0
        0.0
        261119.0
        146219.0

        PAIRS reads detailed summary

        Cis-to-trans ratio is computed as the ratio of long-range cis reads (>20kb) to trans reads plus long-range cis reads. Typically, cis-to-trans ratio higher than 40% is required. Percentage of long-range cis reads is the ratio of long-range cis reads to total number of reads. Minimum 15% is required and 40% or higher suggests a good library(doi:10.1016/j.cell.2014.11.021). Convergence is determined as standard deviation of proportions of four read orientations to be <0.002 (Very good) or <0.05 (Good) (See section Proportion of read orientation versus genomic separation in the pairsqc_report file under pairs/QC/). The slope of log10 contact probability vs distance between 10kb ~ 300kb representing TAD is also provided as well. (See section Contact probability versus genomic separation in the pairsqc_report file under pairs/QC/)

        Showing 4/4 rows and 8/8 columns.
        sampleTotal readsShort cis reads (<20kb)Cis reads (>=20kb)Trans readsCis/Trans ratio% Long-range intrachromosomal readsconvergenceslope
        KD_REP1
        225918.00
        99616.00
        126302.00
        0.00
        100.00
        55.91
        NA
        -0.87
        KD_REP2
        213162.00
        93656.00
        119506.00
        0.00
        100.00
        56.06
        NA
        -0.87
        WT_REP1
        255461.00
        112448.00
        143013.00
        0.00
        100.00
        55.98
        NA
        -0.86
        WT_REP2
        261119.00
        114900.00
        146219.00
        0.00
        100.00
        56.00
        NA
        -0.86

        Proportion of read orientation versus genomic separation

        Contact proportion of reads are shown, stratified by read orientation. Good four cutter samples would show no bias in propotion >3kb(10^3.5).

        loading..

        ATAC R2 reads quality control

        Fraction of reads in peaks (FRiP) – Fraction of all mapped reads that fall into the called peak regions, i.e. usable reads in significantly enriched peaks divided by all usable reads. In general, FRiP scores correlate positively with the number of regions. (Landt et al, Genome Research Sept. 2012, 22(9): 1813–1831). FRiP should be greater than 1%.

        TSS enrichment score is a raio between aggregate distribution of reads centered on TSSs and that flanking the corresponding TSSs. TSS score = the depth of TSS (each 100bp window within 1000 bp each side) / the depth of end flanks (100bp each end). TSSE score = max(mean(TSS score in each window)). TSS enrichment score is calculated according to the definition at ENCODE TSSE score. Transcription start site (TSS) enrichment values are dependent on the reference files used; cutoff values for high quality data are listed in the following table from ENCODE ATAC-seq QC. For HiCAR, only R2 reads is used in TSSE score calculation and reads enrichment is following exponential ratio, we adjusted the TSSE score range. TSSEscore < 2.5: concerning; TSSEscore 2.5-3.5: concerning; TSSEscore > 3.5: Ideal.

        Promoter/Transcript body proportions is test for the R2 reads enriched in promoters or in transcript body.

        Showing 2/2 rows and 5/5 columns.
        Sample NameTSSEscoreFRiPbodyEnrichpromoterEnrichprop.test
        KD
        3.6
        14.2
        5319.0
        6639.0
        0.3
        WT
        4.3
        13.3
        5349.0
        6671.0
        0.3

        MAPS peak calling summary

        Showing 4/4 rows and 20/20 columns.
        runlog10_fdr_cutoffsingleton_fractionAND_sizeAND_mean_distAND_median_distAND_min_countAND_max_pvalAND_max_fdrXOR_sizeXOR_mean_distXOR_median_distXOR_min_countXOR_max_pvalXOR_max_fdrsizemean_distmedian_distmin_countmax_pvalmax_fdr
        KD.10k
        2.0
        0.9
        112.0
        54196.4
        30000.0
        10.0
        0.0
        0.0
        97.0
        41340.2
        20000.0
        12.0
        0.0
        0.0
        209.0
        48229.7
        30000.0
        10.0
        0.0
        0.0
        KD.5k
        2.0
        0.9
        11.0
        29545.5
        10000.0
        12.0
        0.0
        0.0
        32.0
        13750.0
        10000.0
        12.0
        0.0
        0.0
        43.0
        17790.7
        10000.0
        12.0
        0.0
        0.0
        WT.10k
        2.0
        0.8
        184.0
        67391.3
        30000.0
        10.0
        0.0
        0.0
        130.0
        45000.0
        20000.0
        11.0
        0.0
        0.0
        314.0
        58121.0
        30000.0
        10.0
        0.0
        0.0
        WT.5k
        2.0
        0.8
        39.0
        45897.4
        15000.0
        10.0
        0.0
        0.0
        67.0
        16716.4
        10000.0
        11.0
        0.0
        0.0
        106.0
        27452.8
        10000.0
        10.0
        0.0
        0.0

        PCA analysis

        PCA analysis by edgeR.

        loading..

        Peaks genomic element distribution

        The genomic element distribution of the peaks. Promoter region is defined as upstream 2000 and downstream 500 from TSS. geneDownstream is defeined as downstream 2000 from TES.


        The genomeic element distribution for the peaks interacting with promoters

        The genomic element distribution of the other sites of promoter peaks. Promoter region is defined as upstream 2000 and downstream 500 from TSS. geneDownstream is defeined as downstream 2000 from TES.


        Samtools

        Samtools is a suite of programs for interacting with high-throughput sequencing data.

        Samtools Flagstat

        This module parses the output from samtools flagstat. All numbers in millions.

        loading..

        Cutadapt

        Cutadapt is a tool to find and remove adapter sequences, primers, poly-Atails and other types of unwanted sequence from your high-throughput sequencing reads.

        Filtered Reads

        This plot shows the number of reads (SE) / pairs (PE) removed by Cutadapt.

        loading..

        Trimmed Sequence Lengths

        This plot shows the number of reads with certain lengths of adapter trimmed.

        Obs/Exp shows the raw counts divided by the number expected due to sequencing errors. A defined peak may be related to adapter length.

        See the cutadapt documentation for more information on how these numbers are generated.

        loading..

        FastQC

        FastQC is a quality control tool for high throughput sequence data, written by Simon Andrews at the Babraham Institute in Cambridge.

        Sequence Counts

        Sequence counts for each sample. Duplicate read counts are an estimate only.

        This plot show the total number of reads, broken down into unique and duplicate if possible (only more recent versions of FastQC give duplicate info).

        You can read more about duplicate calculation in the FastQC documentation. A small part has been copied here for convenience:

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        loading..

        Sequence Quality Histograms

        The mean quality value across each base position in the read.

        To enable multiple samples to be plotted on the same graph, only the mean quality scores are plotted (unlike the box plots seen in FastQC reports).

        Taken from the FastQC help:

        The y-axis on the graph shows the quality scores. The higher the score, the better the base call. The background of the graph divides the y axis into very good quality calls (green), calls of reasonable quality (orange), and calls of poor quality (red). The quality of calls on most platforms will degrade as the run progresses, so it is common to see base calls falling into the orange area towards the end of a read.

        loading..

        Per Sequence Quality Scores

        The number of reads with average quality scores. Shows if a subset of reads has poor quality.

        From the FastQC help:

        The per sequence quality score report allows you to see if a subset of your sequences have universally low quality values. It is often the case that a subset of sequences will have universally poor quality, however these should represent only a small percentage of the total sequences.

        loading..

        Per Base Sequence Content

        The proportion of each base position for which each of the four normal DNA bases has been called.

        To enable multiple samples to be shown in a single plot, the base composition data is shown as a heatmap. The colours represent the balance between the four bases: an even distribution should give an even muddy brown colour. Hover over the plot to see the percentage of the four bases under the cursor.

        To see the data as a line plot, as in the original FastQC graph, click on a sample track.

        From the FastQC help:

        Per Base Sequence Content plots out the proportion of each base position in a file for which each of the four normal DNA bases has been called.

        In a random library you would expect that there would be little to no difference between the different bases of a sequence run, so the lines in this plot should run parallel with each other. The relative amount of each base should reflect the overall amount of these bases in your genome, but in any case they should not be hugely imbalanced from each other.

        It's worth noting that some types of library will always produce biased sequence composition, normally at the start of the read. Libraries produced by priming using random hexamers (including nearly all RNA-Seq libraries) and those which were fragmented using transposases inherit an intrinsic bias in the positions at which reads start. This bias does not concern an absolute sequence, but instead provides enrichement of a number of different K-mers at the 5' end of the reads. Whilst this is a true technical bias, it isn't something which can be corrected by trimming and in most cases doesn't seem to adversely affect the downstream analysis.

        Click a sample row to see a line plot for that dataset.
        Rollover for sample name
        Position: -
        %T: -
        %C: -
        %A: -
        %G: -

        Per Sequence GC Content

        The average GC content of reads. Normal random library typically have a roughly normal distribution of GC content.

        From the FastQC help:

        This module measures the GC content across the whole length of each sequence in a file and compares it to a modelled normal distribution of GC content.

        In a normal random library you would expect to see a roughly normal distribution of GC content where the central peak corresponds to the overall GC content of the underlying genome. Since we don't know the the GC content of the genome the modal GC content is calculated from the observed data and used to build a reference distribution.

        An unusually shaped distribution could indicate a contaminated library or some other kinds of biased subset. A normal distribution which is shifted indicates some systematic bias which is independent of base position. If there is a systematic bias which creates a shifted normal distribution then this won't be flagged as an error by the module since it doesn't know what your genome's GC content should be.

        loading..

        Per Base N Content

        The percentage of base calls at each position for which an N was called.

        From the FastQC help:

        If a sequencer is unable to make a base call with sufficient confidence then it will normally substitute an N rather than a conventional base call. This graph shows the percentage of base calls at each position for which an N was called.

        It's not unusual to see a very low proportion of Ns appearing in a sequence, especially nearer the end of a sequence. However, if this proportion rises above a few percent it suggests that the analysis pipeline was unable to interpret the data well enough to make valid base calls.

        loading..

        Sequence Length Distribution

        The distribution of fragment sizes (read lengths) found. See the FastQC help

        loading..

        Sequence Duplication Levels

        The relative level of duplication found for every sequence.

        From the FastQC Help:

        In a diverse library most sequences will occur only once in the final set. A low level of duplication may indicate a very high level of coverage of the target sequence, but a high level of duplication is more likely to indicate some kind of enrichment bias (eg PCR over amplification). This graph shows the degree of duplication for every sequence in a library: the relative number of sequences with different degrees of duplication.

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        In a properly diverse library most sequences should fall into the far left of the plot in both the red and blue lines. A general level of enrichment, indicating broad oversequencing in the library will tend to flatten the lines, lowering the low end and generally raising other categories. More specific enrichments of subsets, or the presence of low complexity contaminants will tend to produce spikes towards the right of the plot.

        loading..

        Overrepresented sequences

        The total amount of overrepresented sequences found in each library.

        FastQC calculates and lists overrepresented sequences in FastQ files. It would not be possible to show this for all samples in a MultiQC report, so instead this plot shows the number of sequences categorized as over represented.

        Sometimes, a single sequence may account for a large number of reads in a dataset. To show this, the bars are split into two: the first shows the overrepresented reads that come from the single most common sequence. The second shows the total count from all remaining overrepresented sequences.

        From the FastQC Help:

        A normal high-throughput library will contain a diverse set of sequences, with no individual sequence making up a tiny fraction of the whole. Finding that a single sequence is very overrepresented in the set either means that it is highly biologically significant, or indicates that the library is contaminated, or not as diverse as you expected.

        FastQC lists all of the sequences which make up more than 0.1% of the total. To conserve memory only sequences which appear in the first 100,000 sequences are tracked to the end of the file. It is therefore possible that a sequence which is overrepresented but doesn't appear at the start of the file for some reason could be missed by this module.

        16 samples had less than 1% of reads made up of overrepresented sequences

        Adapter Content

        The cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position.

        Note that only samples with ≥ 0.1% adapter contamination are shown.

        There may be several lines per sample, as one is shown for each adapter detected in the file.

        From the FastQC Help:

        The plot shows a cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position. Once a sequence has been seen in a read it is counted as being present right through to the end of the read so the percentages you see will only increase as the read length goes on.

        loading..

        Status Checks

        Status for each FastQC section showing whether results seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        FastQC assigns a status for each section of the report. These give a quick evaluation of whether the results of the analysis seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        It is important to stress that although the analysis results appear to give a pass/fail result, these evaluations must be taken in the context of what you expect from your library. A 'normal' sample as far as FastQC is concerned is random and diverse. Some experiments may be expected to produce libraries which are biased in particular ways. You should treat the summary evaluations therefore as pointers to where you should concentrate your attention and understand why your library may not look random and diverse.

        Specific guidance on how to interpret the output of each module can be found in the relevant report section, or in the FastQC help.

        In this heatmap, we summarise all of these into a single heatmap for a quick overview. Note that not all FastQC sections have plots in MultiQC reports, but all status checks are shown in this heatmap.

        loading..

        nf-core/hicar Software Versions

        are collected at run time from the software output.

        ATACseqQC
        1.16.0
        ChIPpeakAnno
        3.24.1
        GenomeInfoDb
        1.26.4
        GenomicFeatures
        1.44.0
        MAPS
        1.1.0
        awk
        5.1.0
        bedtools
        2.30.0
        bwa
        0.7.17-r1188
        cooler
        0.8.11
        cutadapt
        3.4
        edgeR
        3.32.1
        fastqc
        0.11.9
        gzip
        321.100.11
        macs2
        2.2.7.1
        nextflow
        21.05.0.edge
        nf-core/hicar
        1.0dev
        pairtools
        0.3.0
        perl
        5.026002
        rtracklayer
        1.50.0
        samtools
        1.13
        ucsc
        2

        nf-core/hicar Workflow Summary

        - this information is collected when the pipeline is started.

        Core Nextflow options

        runName
        mighty_boyd
        launchDir
        /Users/ouj/github/new
        workDir
        /Users/ouj/github/new/work
        projectDir
        /Users/ouj/github/new/nf-core-hicar
        userName
        ouj
        profile
        conda,test_full
        configFiles
        /Users/ouj/github/new/nf-core-hicar/nextflow.config

        Input/output options

        input
        https://raw.githubusercontent.com/jianhong/nf-core-hicar/master/assets/samplesheet.csv
        outdir
        full_test_results

        Reference genome options

        fasta
        https://raw.githubusercontent.com/nf-core/test-datasets/hicar/data/genomics/homo_sapiens/fasta/chr22.fa.gz
        gtf
        https://raw.githubusercontent.com/nf-core/test-datasets/hicar/data/genomics/homo_sapiens/annotation/chr22.gtf.gz
        mappability
        https://raw.githubusercontent.com/nf-core/test-datasets/hicar/data/genomics/homo_sapiens/mappability/wgEncodeCrgMapabilityAlign50mer.chr22.bigWig
        macs_gsize
        3e7
        ucscname
        hg19

        MACS2 peak calling options

        qval_thresh
        0.05

        MAPS peak calling options

        maps_cutoff_counts
        6

        Other options not expose

        enrichment_fdr
        1

        Institutional config options

        config_profile_name
        Full test profile
        config_profile_description
        Full test dataset to check pipeline function

        Max job request options

        max_cpus
        2
        max_memory
        6 GB
        max_time
        6h

        Generic options

        tracedir
        ./results/pipeline_info
        enable_conda
        true