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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 2024-03-28, 01:33 based on data in: /vast/gencore/GENEFLOW/work/0f/2e06c6382bbeb9335e808d39f277d0/merged


        General Statistics

        Showing 160/160 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HWTKNDRX3_n01_10G_1-17_NEB
        79.1%
        48%
        37.2
        HWTKNDRX3_n01_10R_1-17_NEB
        85.2%
        50%
        22.0
        HWTKNDRX3_n01_11R_1-17_NEB
        82.1%
        50%
        26.8
        HWTKNDRX3_n01_12G_1-17_NEB
        80.4%
        48%
        46.9
        HWTKNDRX3_n01_12R_1-17_NEB
        78.7%
        49%
        31.1
        HWTKNDRX3_n01_13G_1-17_NEB
        76.5%
        48%
        29.8
        HWTKNDRX3_n01_13R_1-17_NEB
        76.9%
        49%
        29.3
        HWTKNDRX3_n01_14G_1-17_NEB
        79.8%
        49%
        30.6
        HWTKNDRX3_n01_14R_1-17_NEB
        77.3%
        49%
        24.6
        HWTKNDRX3_n01_15G_1-17_NEB
        76.6%
        48%
        31.4
        HWTKNDRX3_n01_15R_1-17_NEB
        81.2%
        50%
        26.1
        HWTKNDRX3_n01_1R_1-17_NEB
        86.2%
        50%
        19.7
        HWTKNDRX3_n01_2G_1-17_NEB
        80.0%
        48%
        25.0
        HWTKNDRX3_n01_3G_1-17_NEB
        81.5%
        50%
        24.1
        HWTKNDRX3_n01_3R_1-17_NEB
        83.3%
        50%
        21.2
        HWTKNDRX3_n01_4G_1-17_NEB
        83.3%
        51%
        29.8
        HWTKNDRX3_n01_4R_1-17_NEB
        82.8%
        49%
        22.1
        HWTKNDRX3_n01_5G_1-17_NEB
        74.5%
        48%
        27.2
        HWTKNDRX3_n01_5R_1-17_NEB
        86.1%
        50%
        22.3
        HWTKNDRX3_n01_6R_1-17_NEB
        85.4%
        50%
        19.3
        HWTKNDRX3_n01_7G_1-17_NEB
        80.8%
        47%
        24.8
        HWTKNDRX3_n01_7R_1-17_NEB
        86.7%
        49%
        19.6
        HWTKNDRX3_n01_8G_1-17_NEB
        75.0%
        48%
        24.5
        HWTKNDRX3_n01_8R_1-17_NEB
        88.3%
        51%
        22.5
        HWTKNDRX3_n01_9G_1-17_NEB
        81.5%
        50%
        28.7
        HWTKNDRX3_n01_9R_1-17_NEB
        77.5%
        49%
        20.9
        HWTKNDRX3_n01_AH_01
        85.1%
        48%
        22.6
        HWTKNDRX3_n01_AH_02
        82.4%
        48%
        23.0
        HWTKNDRX3_n01_AH_03
        83.4%
        46%
        4.3
        HWTKNDRX3_n01_AH_04
        81.1%
        47%
        17.2
        HWTKNDRX3_n01_AH_05
        85.2%
        46%
        9.9
        HWTKNDRX3_n01_AH_06
        74.7%
        48%
        16.7
        HWTKNDRX3_n01_AH_07
        83.0%
        48%
        15.4
        HWTKNDRX3_n01_AH_08
        83.2%
        49%
        7.5
        HWTKNDRX3_n01_AH_09
        83.8%
        49%
        24.6
        HWTKNDRX3_n01_AH_10
        88.0%
        49%
        17.8
        HWTKNDRX3_n01_AH_11
        83.7%
        46%
        24.7
        HWTKNDRX3_n01_AH_12
        82.4%
        47%
        17.2
        HWTKNDRX3_n01_AH_13
        87.7%
        47%
        12.5
        HWTKNDRX3_n01_AH_14
        86.3%
        47%
        11.1
        HWTKNDRX3_n01_AH_15
        85.1%
        49%
        13.6
        HWTKNDRX3_n01_AH_16
        82.5%
        49%
        7.0
        HWTKNDRX3_n01_AH_17
        87.5%
        48%
        14.6
        HWTKNDRX3_n01_AH_18
        84.8%
        48%
        12.8
        HWTKNDRX3_n01_AJ_1
        82.8%
        55%
        21.4
        HWTKNDRX3_n01_AJ_2
        85.0%
        38%
        13.1
        HWTKNDRX3_n01_AJ_3
        80.4%
        51%
        18.1
        HWTKNDRX3_n01_AJ_4
        66.1%
        52%
        10.0
        HWTKNDRX3_n01_AJ_5
        70.5%
        52%
        13.4
        HWTKNDRX3_n01_AJ_6
        75.3%
        50%
        13.7
        HWTKNDRX3_n01_AJ_7
        77.5%
        47%
        15.9
        HWTKNDRX3_n01_AJ_8
        80.7%
        46%
        21.0
        HWTKNDRX3_n01_LA_1
        78.4%
        47%
        27.6
        HWTKNDRX3_n01_LA_10
        85.4%
        49%
        36.0
        HWTKNDRX3_n01_LA_11
        75.0%
        49%
        24.6
        HWTKNDRX3_n01_LA_12
        91.9%
        52%
        83.0
        HWTKNDRX3_n01_LA_13
        73.4%
        48%
        16.4
        HWTKNDRX3_n01_LA_15
        82.5%
        49%
        33.9
        HWTKNDRX3_n01_LA_16
        75.9%
        48%
        22.6
        HWTKNDRX3_n01_LA_17
        77.0%
        48%
        23.8
        HWTKNDRX3_n01_LA_18
        79.9%
        48%
        45.3
        HWTKNDRX3_n01_LA_19
        79.3%
        48%
        25.6
        HWTKNDRX3_n01_LA_2
        82.5%
        46%
        32.4
        HWTKNDRX3_n01_LA_20
        78.9%
        47%
        30.3
        HWTKNDRX3_n01_LA_21
        80.0%
        48%
        23.2
        HWTKNDRX3_n01_LA_22
        82.8%
        46%
        64.5
        HWTKNDRX3_n01_LA_23
        75.8%
        46%
        23.3
        HWTKNDRX3_n01_LA_24
        94.8%
        54%
        42.8
        HWTKNDRX3_n01_LA_25
        78.3%
        47%
        34.4
        HWTKNDRX3_n01_LA_26
        83.7%
        46%
        65.8
        HWTKNDRX3_n01_LA_27
        82.7%
        46%
        67.1
        HWTKNDRX3_n01_LA_28
        80.2%
        46%
        40.5
        HWTKNDRX3_n01_LA_29
        83.1%
        47%
        28.4
        HWTKNDRX3_n01_LA_3
        80.9%
        48%
        28.1
        HWTKNDRX3_n01_LA_30
        77.3%
        48%
        23.3
        HWTKNDRX3_n01_LA_5
        79.4%
        48%
        28.0
        HWTKNDRX3_n01_LA_6
        80.9%
        48%
        24.3
        HWTKNDRX3_n01_LA_7
        80.6%
        50%
        28.4
        HWTKNDRX3_n01_LA_8
        77.2%
        47%
        27.3
        HWTKNDRX3_n01_undetermined
        74.3%
        44%
        70.7
        HWTKNDRX3_n02_10G_1-17_NEB
        77.3%
        48%
        37.2
        HWTKNDRX3_n02_10R_1-17_NEB
        82.6%
        51%
        22.0
        HWTKNDRX3_n02_11R_1-17_NEB
        79.6%
        50%
        26.8
        HWTKNDRX3_n02_12G_1-17_NEB
        79.0%
        48%
        46.9
        HWTKNDRX3_n02_12R_1-17_NEB
        76.9%
        49%
        31.1
        HWTKNDRX3_n02_13G_1-17_NEB
        74.0%
        48%
        29.8
        HWTKNDRX3_n02_13R_1-17_NEB
        75.3%
        49%
        29.3
        HWTKNDRX3_n02_14G_1-17_NEB
        78.0%
        49%
        30.6
        HWTKNDRX3_n02_14R_1-17_NEB
        75.4%
        49%
        24.6
        HWTKNDRX3_n02_15G_1-17_NEB
        75.3%
        48%
        31.4
        HWTKNDRX3_n02_15R_1-17_NEB
        79.1%
        50%
        26.1
        HWTKNDRX3_n02_1R_1-17_NEB
        83.9%
        51%
        19.7
        HWTKNDRX3_n02_2G_1-17_NEB
        78.0%
        48%
        25.0
        HWTKNDRX3_n02_3G_1-17_NEB
        78.8%
        51%
        24.1
        HWTKNDRX3_n02_3R_1-17_NEB
        81.8%
        50%
        21.2
        HWTKNDRX3_n02_4G_1-17_NEB
        80.4%
        52%
        29.8
        HWTKNDRX3_n02_4R_1-17_NEB
        81.1%
        49%
        22.1
        HWTKNDRX3_n02_5G_1-17_NEB
        72.5%
        48%
        27.2
        HWTKNDRX3_n02_5R_1-17_NEB
        83.5%
        50%
        22.3
        HWTKNDRX3_n02_6R_1-17_NEB
        82.5%
        52%
        19.3
        HWTKNDRX3_n02_7G_1-17_NEB
        78.8%
        48%
        24.8
        HWTKNDRX3_n02_7R_1-17_NEB
        84.3%
        50%
        19.6
        HWTKNDRX3_n02_8G_1-17_NEB
        73.2%
        48%
        24.5
        HWTKNDRX3_n02_8R_1-17_NEB
        84.0%
        52%
        22.5
        HWTKNDRX3_n02_9G_1-17_NEB
        77.5%
        50%
        28.7
        HWTKNDRX3_n02_9R_1-17_NEB
        75.4%
        49%
        20.9
        HWTKNDRX3_n02_AH_01
        83.2%
        48%
        22.6
        HWTKNDRX3_n02_AH_02
        80.5%
        48%
        23.0
        HWTKNDRX3_n02_AH_03
        81.9%
        47%
        4.3
        HWTKNDRX3_n02_AH_04
        79.1%
        47%
        17.2
        HWTKNDRX3_n02_AH_05
        83.7%
        46%
        9.9
        HWTKNDRX3_n02_AH_06
        73.3%
        48%
        16.7
        HWTKNDRX3_n02_AH_07
        81.4%
        48%
        15.4
        HWTKNDRX3_n02_AH_08
        81.8%
        50%
        7.5
        HWTKNDRX3_n02_AH_09
        82.4%
        49%
        24.6
        HWTKNDRX3_n02_AH_10
        87.0%
        49%
        17.8
        HWTKNDRX3_n02_AH_11
        81.0%
        46%
        24.7
        HWTKNDRX3_n02_AH_12
        80.1%
        47%
        17.2
        HWTKNDRX3_n02_AH_13
        86.1%
        47%
        12.5
        HWTKNDRX3_n02_AH_14
        84.9%
        47%
        11.1
        HWTKNDRX3_n02_AH_15
        83.3%
        49%
        13.6
        HWTKNDRX3_n02_AH_16
        81.1%
        49%
        7.0
        HWTKNDRX3_n02_AH_17
        85.5%
        48%
        14.6
        HWTKNDRX3_n02_AH_18
        82.9%
        48%
        12.8
        HWTKNDRX3_n02_AJ_1
        73.6%
        61%
        21.4
        HWTKNDRX3_n02_AJ_2
        82.5%
        40%
        13.1
        HWTKNDRX3_n02_AJ_3
        75.5%
        54%
        18.1
        HWTKNDRX3_n02_AJ_4
        59.8%
        55%
        10.0
        HWTKNDRX3_n02_AJ_5
        64.1%
        55%
        13.4
        HWTKNDRX3_n02_AJ_6
        70.7%
        52%
        13.7
        HWTKNDRX3_n02_AJ_7
        73.3%
        48%
        15.9
        HWTKNDRX3_n02_AJ_8
        77.8%
        47%
        21.0
        HWTKNDRX3_n02_LA_1
        77.2%
        48%
        27.6
        HWTKNDRX3_n02_LA_10
        82.5%
        50%
        36.0
        HWTKNDRX3_n02_LA_11
        73.1%
        50%
        24.6
        HWTKNDRX3_n02_LA_12
        90.2%
        52%
        83.0
        HWTKNDRX3_n02_LA_13
        71.3%
        49%
        16.4
        HWTKNDRX3_n02_LA_15
        79.6%
        50%
        33.9
        HWTKNDRX3_n02_LA_16
        73.7%
        48%
        22.6
        HWTKNDRX3_n02_LA_17
        75.3%
        48%
        23.8
        HWTKNDRX3_n02_LA_18
        77.8%
        49%
        45.3
        HWTKNDRX3_n02_LA_19
        77.5%
        48%
        25.6
        HWTKNDRX3_n02_LA_2
        81.2%
        46%
        32.4
        HWTKNDRX3_n02_LA_20
        77.3%
        47%
        30.3
        HWTKNDRX3_n02_LA_21
        77.8%
        48%
        23.2
        HWTKNDRX3_n02_LA_22
        80.8%
        46%
        64.5
        HWTKNDRX3_n02_LA_23
        73.7%
        46%
        23.3
        HWTKNDRX3_n02_LA_24
        93.5%
        54%
        42.8
        HWTKNDRX3_n02_LA_25
        75.4%
        47%
        34.4
        HWTKNDRX3_n02_LA_26
        82.1%
        46%
        65.8
        HWTKNDRX3_n02_LA_27
        79.3%
        46%
        67.1
        HWTKNDRX3_n02_LA_28
        78.5%
        46%
        40.5
        HWTKNDRX3_n02_LA_29
        81.1%
        47%
        28.4
        HWTKNDRX3_n02_LA_3
        79.0%
        48%
        28.1
        HWTKNDRX3_n02_LA_30
        75.6%
        48%
        23.3
        HWTKNDRX3_n02_LA_5
        77.2%
        48%
        28.0
        HWTKNDRX3_n02_LA_6
        79.2%
        49%
        24.3
        HWTKNDRX3_n02_LA_7
        78.1%
        50%
        28.4
        HWTKNDRX3_n02_LA_8
        75.3%
        48%
        27.3
        HWTKNDRX3_n02_undetermined
        68.8%
        44%
        70.7

        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 80/80 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        70721467
        3.4
        1R_1-17_NEB
        19731223
        0.9
        3R_1-17_NEB
        21190437
        1.0
        4R_1-17_NEB
        22110584
        1.0
        5R_1-17_NEB
        22296319
        1.1
        6R_1-17_NEB
        19281673
        0.9
        7R_1-17_NEB
        19584598
        0.9
        8R_1-17_NEB
        22475506
        1.1
        9R_1-17_NEB
        20937586
        1.0
        10R_1-17_NEB
        21957238
        1.0
        11R_1-17_NEB
        26800804
        1.3
        12R_1-17_NEB
        31097681
        1.5
        13R_1-17_NEB
        29317332
        1.4
        14R_1-17_NEB
        24580620
        1.2
        15R_1-17_NEB
        26092814
        1.2
        2G_1-17_NEB
        24963856
        1.2
        3G_1-17_NEB
        24121638
        1.1
        4G_1-17_NEB
        29791457
        1.4
        5G_1-17_NEB
        27161675
        1.3
        7G_1-17_NEB
        24775543
        1.2
        8G_1-17_NEB
        24515341
        1.2
        9G_1-17_NEB
        28740005
        1.4
        10G_1-17_NEB
        37162521
        1.8
        12G_1-17_NEB
        46939087
        2.2
        13G_1-17_NEB
        29844737
        1.4
        14G_1-17_NEB
        30566269
        1.5
        15G_1-17_NEB
        31404525
        1.5
        AH_01
        22625368
        1.1
        AH_02
        22998315
        1.1
        AH_03
        4305714
        0.2
        AH_04
        17204439
        0.8
        AH_05
        9881004
        0.5
        AH_06
        16670175
        0.8
        AH_07
        15427218
        0.7
        AH_08
        7469394
        0.4
        AH_09
        24594282
        1.2
        AH_10
        17803840
        0.8
        AH_11
        24697882
        1.2
        AH_12
        17233968
        0.8
        AH_13
        12500229
        0.6
        AH_14
        11052077
        0.5
        AH_15
        13641202
        0.6
        AH_16
        7040405
        0.3
        AH_17
        14628722
        0.7
        AH_18
        12801529
        0.6
        AJ_1
        21397001
        1.0
        AJ_2
        13100295
        0.6
        AJ_3
        18116984
        0.9
        AJ_4
        10049407
        0.5
        AJ_5
        13405373
        0.6
        AJ_6
        13657100
        0.6
        AJ_7
        15922182
        0.8
        AJ_8
        20984960
        1.0
        LA_1
        27559498
        1.3
        LA_2
        32350624
        1.5
        LA_3
        28057073
        1.3
        LA_5
        28042767
        1.3
        LA_6
        24344136
        1.2
        LA_7
        28413840
        1.3
        LA_8
        27309688
        1.3
        LA_10
        36037376
        1.7
        LA_11
        24579251
        1.2
        LA_12
        83034542
        3.9
        LA_13
        16431898
        0.8
        LA_15
        33864146
        1.6
        LA_16
        22619979
        1.1
        LA_17
        23782056
        1.1
        LA_18
        45308740
        2.1
        LA_19
        25647023
        1.2
        LA_20
        30283206
        1.4
        LA_21
        23154252
        1.1
        LA_22
        64516678
        3.1
        LA_23
        23295742
        1.1
        LA_24
        42802887
        2.0
        LA_25
        34416483
        1.6
        LA_26
        65820669
        3.1
        LA_27
        67073461
        3.2
        LA_28
        40460110
        1.9
        LA_29
        28358843
        1.3
        LA_30
        23261491
        1.1

        Barcodes of Undetermined Reads


        We have determined the barcodes of your undetermined reads. Here are the top 20 barcodes. The full list is available here. If your libraries are dual indexed, the two indices are concatenated.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGGGG
        46015702.0
        65.1
        AGTGACCT
        123110.0
        0.2
        CAAGGTTA
        84412.0
        0.1
        CGGGGGGG
        70167.0
        0.1
        CAACAAAA
        69494.0
        0.1
        CTTCCATA
        67810.0
        0.1
        GGGGGGGC
        67546.0
        0.1
        CAAAAAAA
        66409.0
        0.1
        GCCTTACA
        65217.0
        0.1
        GCCTAACA
        62712.0
        0.1
        GGGGGGTG
        58651.0
        0.1
        AACAAAAC
        57165.0
        0.1
        CCAGGTTA
        52883.0
        0.1
        ACGCAATA
        52165.0
        0.1
        GGGGGGGT
        49958.0
        0.1
        CACCAAAA
        46731.0
        0.1
        CCTTCATA
        45670.0
        0.1
        CCTCCATA
        44496.0
        0.1
        AACAAAAA
        42444.0
        0.1
        GCAATAAC
        41977.0
        0.1

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        2.0
        2553348096
        2108198060
        3.4
        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 (101bp).

        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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