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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 1.9

        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.

        Report generated on 2023-02-05, 18:34 based on data in: /scratch/gencore/logs/html/HN5YJDRX2/merged


        General Statistics

        Showing 130/130 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HN5YJDRX2_n01_br248a
        74.6%
        50%
        1.9
        HN5YJDRX2_n01_br249a
        43.1%
        54%
        13.0
        HN5YJDRX2_n01_br250a
        24.6%
        52%
        6.6
        HN5YJDRX2_n01_br251a
        52.3%
        62%
        7.3
        HN5YJDRX2_n01_br252a
        69.0%
        47%
        3.4
        HN5YJDRX2_n01_br253a
        27.5%
        54%
        9.7
        HN5YJDRX2_n01_br254a
        38.7%
        52%
        18.5
        HN5YJDRX2_n01_br255a
        70.8%
        62%
        16.7
        HN5YJDRX2_n01_br256a
        73.1%
        51%
        4.2
        HN5YJDRX2_n01_br257a
        42.4%
        53%
        19.3
        HN5YJDRX2_n01_br258a
        15.7%
        50%
        9.4
        HN5YJDRX2_n01_br259a
        72.2%
        62%
        21.5
        HN5YJDRX2_n01_br260a
        80.3%
        48%
        2.6
        HN5YJDRX2_n01_br261a
        55.0%
        54%
        17.5
        HN5YJDRX2_n01_br262a
        46.3%
        50%
        37.5
        HN5YJDRX2_n01_br263a
        71.2%
        63%
        20.1
        HN5YJDRX2_n01_br264a
        57.4%
        47%
        10.0
        HN5YJDRX2_n01_br265a
        50.1%
        55%
        24.9
        HN5YJDRX2_n01_br266a
        23.0%
        52%
        28.4
        HN5YJDRX2_n01_br267a
        53.9%
        64%
        6.2
        HN5YJDRX2_n01_br268a
        74.9%
        49%
        3.1
        HN5YJDRX2_n01_br269a
        41.3%
        55%
        11.9
        HN5YJDRX2_n01_br270a
        13.2%
        51%
        10.3
        HN5YJDRX2_n01_br271a
        66.8%
        63%
        22.1
        HN5YJDRX2_n01_br272a
        86.1%
        44%
        4.3
        HN5YJDRX2_n01_br273a
        62.0%
        57%
        15.5
        HN5YJDRX2_n01_br274a
        36.2%
        54%
        9.5
        HN5YJDRX2_n01_br275a
        63.9%
        63%
        9.5
        HN5YJDRX2_n01_br276a
        88.5%
        45%
        4.2
        HN5YJDRX2_n01_br277a
        60.3%
        56%
        14.6
        HN5YJDRX2_n01_br278a
        32.1%
        51%
        15.1
        HN5YJDRX2_n01_br279a
        75.8%
        63%
        23.6
        HN5YJDRX2_n01_br280a
        73.3%
        47%
        2.0
        HN5YJDRX2_n01_br281a
        57.3%
        53%
        27.2
        HN5YJDRX2_n01_br282a
        25.6%
        52%
        14.9
        HN5YJDRX2_n01_br283a
        71.9%
        62%
        23.2
        HN5YJDRX2_n01_br284a
        80.1%
        47%
        4.2
        HN5YJDRX2_n01_br285a
        36.2%
        54%
        10.9
        HN5YJDRX2_n01_br286a
        41.3%
        52%
        39.2
        HN5YJDRX2_n01_br287a
        66.9%
        62%
        18.7
        HN5YJDRX2_n01_br288a
        82.1%
        48%
        3.0
        HN5YJDRX2_n01_br289a
        52.3%
        56%
        14.0
        HN5YJDRX2_n01_br290a
        41.6%
        51%
        23.0
        HN5YJDRX2_n01_br291a
        67.4%
        63%
        19.1
        HN5YJDRX2_n01_br292a
        81.3%
        48%
        2.6
        HN5YJDRX2_n01_br293a
        81.6%
        55%
        48.4
        HN5YJDRX2_n01_br294a
        38.3%
        51%
        21.4
        HN5YJDRX2_n01_br295a
        74.9%
        63%
        28.8
        HN5YJDRX2_n01_br296a
        67.2%
        46%
        1.8
        HN5YJDRX2_n01_br297a
        46.8%
        58%
        6.5
        HN5YJDRX2_n01_br298a
        28.1%
        51%
        15.2
        HN5YJDRX2_n01_br299a
        59.7%
        63%
        11.0
        HN5YJDRX2_n01_br300a
        65.4%
        48%
        0.7
        HN5YJDRX2_n01_br301a
        60.0%
        57%
        10.4
        HN5YJDRX2_n01_br302a
        49.8%
        49%
        20.9
        HN5YJDRX2_n01_br303a
        66.0%
        64%
        15.7
        HN5YJDRX2_n01_br304a
        83.0%
        51%
        2.8
        HN5YJDRX2_n01_br305a
        55.3%
        59%
        8.4
        HN5YJDRX2_n01_br306a
        38.5%
        55%
        10.0
        HN5YJDRX2_n01_br307a
        71.8%
        63%
        20.2
        HN5YJDRX2_n01_br308a
        85.0%
        47%
        2.8
        HN5YJDRX2_n01_br309a
        49.0%
        59%
        6.2
        HN5YJDRX2_n01_br310a
        34.1%
        51%
        20.9
        HN5YJDRX2_n01_br311a
        73.2%
        63%
        26.1
        HN5YJDRX2_n01_undetermined
        45.3%
        50%
        63.4
        HN5YJDRX2_n02_br248a
        73.6%
        50%
        1.9
        HN5YJDRX2_n02_br249a
        41.4%
        54%
        13.0
        HN5YJDRX2_n02_br250a
        23.6%
        52%
        6.6
        HN5YJDRX2_n02_br251a
        48.4%
        61%
        7.3
        HN5YJDRX2_n02_br252a
        64.6%
        47%
        3.4
        HN5YJDRX2_n02_br253a
        25.7%
        54%
        9.7
        HN5YJDRX2_n02_br254a
        37.7%
        52%
        18.5
        HN5YJDRX2_n02_br255a
        67.2%
        62%
        16.7
        HN5YJDRX2_n02_br256a
        72.7%
        50%
        4.2
        HN5YJDRX2_n02_br257a
        40.0%
        53%
        19.3
        HN5YJDRX2_n02_br258a
        15.1%
        50%
        9.4
        HN5YJDRX2_n02_br259a
        66.7%
        62%
        21.5
        HN5YJDRX2_n02_br260a
        76.1%
        47%
        2.6
        HN5YJDRX2_n02_br261a
        50.0%
        53%
        17.5
        HN5YJDRX2_n02_br262a
        45.0%
        49%
        37.5
        HN5YJDRX2_n02_br263a
        65.3%
        62%
        20.1
        HN5YJDRX2_n02_br264a
        63.2%
        47%
        10.0
        HN5YJDRX2_n02_br265a
        53.9%
        54%
        24.9
        HN5YJDRX2_n02_br266a
        28.1%
        51%
        28.4
        HN5YJDRX2_n02_br267a
        59.7%
        64%
        6.2
        HN5YJDRX2_n02_br268a
        81.3%
        48%
        3.1
        HN5YJDRX2_n02_br269a
        44.5%
        55%
        11.9
        HN5YJDRX2_n02_br270a
        16.0%
        51%
        10.3
        HN5YJDRX2_n02_br271a
        71.8%
        62%
        22.1
        HN5YJDRX2_n02_br272a
        82.4%
        44%
        4.3
        HN5YJDRX2_n02_br273a
        58.2%
        57%
        15.5
        HN5YJDRX2_n02_br274a
        34.9%
        53%
        9.5
        HN5YJDRX2_n02_br275a
        61.1%
        63%
        9.5
        HN5YJDRX2_n02_br276a
        83.8%
        45%
        4.2
        HN5YJDRX2_n02_br277a
        54.9%
        56%
        14.6
        HN5YJDRX2_n02_br278a
        31.2%
        51%
        15.1
        HN5YJDRX2_n02_br279a
        70.7%
        63%
        23.6
        HN5YJDRX2_n02_br280a
        70.9%
        46%
        2.0
        HN5YJDRX2_n02_br281a
        53.9%
        53%
        27.2
        HN5YJDRX2_n02_br282a
        24.3%
        52%
        14.9
        HN5YJDRX2_n02_br283a
        66.6%
        62%
        23.2
        HN5YJDRX2_n02_br284a
        75.1%
        46%
        4.2
        HN5YJDRX2_n02_br285a
        32.5%
        53%
        10.9
        HN5YJDRX2_n02_br286a
        39.8%
        52%
        39.2
        HN5YJDRX2_n02_br287a
        61.5%
        62%
        18.7
        HN5YJDRX2_n02_br288a
        80.1%
        47%
        3.0
        HN5YJDRX2_n02_br289a
        50.4%
        55%
        14.0
        HN5YJDRX2_n02_br290a
        39.6%
        50%
        23.0
        HN5YJDRX2_n02_br291a
        66.5%
        63%
        19.1
        HN5YJDRX2_n02_br292a
        77.6%
        47%
        2.6
        HN5YJDRX2_n02_br293a
        74.6%
        55%
        48.4
        HN5YJDRX2_n02_br294a
        37.5%
        50%
        21.4
        HN5YJDRX2_n02_br295a
        69.5%
        63%
        28.8
        HN5YJDRX2_n02_br296a
        68.7%
        46%
        1.8
        HN5YJDRX2_n02_br297a
        48.2%
        57%
        6.5
        HN5YJDRX2_n02_br298a
        29.0%
        50%
        15.2
        HN5YJDRX2_n02_br299a
        61.1%
        63%
        11.0
        HN5YJDRX2_n02_br300a
        66.5%
        47%
        0.7
        HN5YJDRX2_n02_br301a
        57.6%
        57%
        10.4
        HN5YJDRX2_n02_br302a
        51.9%
        49%
        20.9
        HN5YJDRX2_n02_br303a
        68.7%
        63%
        15.7
        HN5YJDRX2_n02_br304a
        82.0%
        50%
        2.8
        HN5YJDRX2_n02_br305a
        53.2%
        59%
        8.4
        HN5YJDRX2_n02_br306a
        37.5%
        54%
        10.0
        HN5YJDRX2_n02_br307a
        67.2%
        63%
        20.2
        HN5YJDRX2_n02_br308a
        82.2%
        47%
        2.8
        HN5YJDRX2_n02_br309a
        46.6%
        58%
        6.2
        HN5YJDRX2_n02_br310a
        33.4%
        51%
        20.9
        HN5YJDRX2_n02_br311a
        68.6%
        63%
        26.1
        HN5YJDRX2_n02_undetermined
        39.7%
        51%
        63.4

        Demultiplexing Report


        Total Read Count: Total number of PF (Passing Filter) reads in this library.
        Portion: The proportion of reads that represent the individual library in the entire Library Pool.

        Showing 65/65 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        63371528
        6.6
        br248a
        1857783
        0.2
        br249a
        13038264
        1.3
        br250a
        6608262
        0.7
        br251a
        7337933
        0.8
        br252a
        3415337
        0.4
        br253a
        9711674
        1.0
        br254a
        18513791
        1.9
        br255a
        16719757
        1.7
        br256a
        4172336
        0.4
        br257a
        19287177
        2.0
        br258a
        9432151
        1.0
        br259a
        21536208
        2.2
        br260a
        2600917
        0.3
        br261a
        17501633
        1.8
        br262a
        37469860
        3.9
        br263a
        20086680
        2.1
        br264a
        10027407
        1.0
        br265a
        24915130
        2.6
        br266a
        28363583
        2.9
        br267a
        6247230
        0.6
        br268a
        3056082
        0.3
        br269a
        11911616
        1.2
        br270a
        10268969
        1.1
        br271a
        22094785
        2.3
        br272a
        4328163
        0.4
        br273a
        15463005
        1.6
        br274a
        9453976
        1.0
        br275a
        9468241
        1.0
        br276a
        4182809
        0.4
        br277a
        14649438
        1.5
        br278a
        15131900
        1.6
        br279a
        23604142
        2.4
        br280a
        1966499
        0.2
        br281a
        27199023
        2.8
        br282a
        14911170
        1.5
        br283a
        23195316
        2.4
        br284a
        4209802
        0.4
        br285a
        10933146
        1.1
        br286a
        39178476
        4.1
        br287a
        18682401
        1.9
        br288a
        2955425
        0.3
        br289a
        13985778
        1.4
        br290a
        23013662
        2.4
        br291a
        19144034
        2.0
        br292a
        2623110
        0.3
        br293a
        48432274
        5.0
        br294a
        21383860
        2.2
        br295a
        28764002
        3.0
        br296a
        1820771
        0.2
        br297a
        6522511
        0.7
        br298a
        15202941
        1.6
        br299a
        10965692
        1.1
        br300a
        703546
        0.1
        br301a
        10395889
        1.1
        br302a
        20851851
        2.2
        br303a
        15704538
        1.6
        br304a
        2764321
        0.3
        br305a
        8440876
        0.9
        br306a
        10012391
        1.0
        br307a
        20167892
        2.1
        br308a
        2833593
        0.3
        br309a
        6204287
        0.6
        br310a
        20912351
        2.2
        br311a
        26133392
        2.7

        Barcodes of Undetermined Reads


        We have determined the barcodes of your undetermined reads (reads containing a barcode that you did not encode in your metadata). Here are the top 20 barcodes belonging to the undetermined reads. The full list is available here. If your libraries are dual indexed, the two indicies are concatenated.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGGGGAGATCTCG
        14406179.0
        22.7
        GGGGGGGGAGCTCTAA
        1333042.0
        2.1
        GGGGGGGGCGTAGCTT
        1077698.0
        1.7
        GGGGGGGGAGTGATTC
        832510.0
        1.3
        GGGGGGGGGAGTTAAG
        826364.0
        1.3
        GGGGGGGGCCGCATGT
        714751.0
        1.1
        GGGGGGGGCATCCACC
        605733.0
        1.0
        GGGGGGGGAGTTCTCG
        589141.0
        0.9
        CCACTCCTGGGGGGGG
        554364.0
        0.9
        GGGGGGGGGGATACCA
        520637.0
        0.8
        GGGGGGGGTAAGGTGG
        493274.0
        0.8
        GGGGGGGGGGTTCTCG
        471760.0
        0.7
        CCACTCCTGGTGGTGG
        451324.0
        0.7
        GGGGGGGGCGATCTCG
        400500.0
        0.6
        GGGGGGGGCGTTCTCG
        393967.0
        0.6
        CTCTCTACGGGGGGGG
        358933.0
        0.6
        CCACTCCTGGGGGTGG
        336975.0
        0.5
        GGGGGGGGGGTGCTCG
        328595.0
        0.5
        GGGGGGGGTGTTCTCG
        319382.0
        0.5
        CTCTCTACGGTGGTGG
        311397.0
        0.5

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        2.0
        1276674048
        966042587
        6.6
        1.9

        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Length Distribution

        All samples have sequences of a single length (151bp).

        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

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