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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 2022-04-09, 21:58 based on data in: /scratch/gencore/logs/html/HC33HDMXY/merged


        General Statistics

        Showing 194/194 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HC33HDMXY_n01_T0_A04
        77.5%
        40%
        52.1
        HC33HDMXY_n01_T0_A06
        75.2%
        40%
        45.5
        HC33HDMXY_n01_T0_A10
        75.8%
        39%
        45.2
        HC33HDMXY_n01_T0_B03
        76.9%
        40%
        51.8
        HC33HDMXY_n01_T0_B09
        76.6%
        40%
        49.9
        HC33HDMXY_n01_T0_C04
        74.0%
        40%
        40.0
        HC33HDMXY_n01_T0_C06
        75.2%
        40%
        43.9
        HC33HDMXY_n01_T0_C10
        78.9%
        39%
        56.6
        HC33HDMXY_n01_T0_D03
        77.6%
        39%
        51.9
        HC33HDMXY_n01_T0_D09
        75.9%
        40%
        45.6
        HC33HDMXY_n01_T0_E04
        71.4%
        40%
        38.8
        HC33HDMXY_n01_T0_E06
        69.0%
        40%
        27.6
        HC33HDMXY_n01_T0_E10
        74.8%
        40%
        41.3
        HC33HDMXY_n01_T0_F03
        72.2%
        40%
        35.2
        HC33HDMXY_n01_T0_F09
        77.1%
        40%
        52.3
        HC33HDMXY_n01_T0_G04
        74.7%
        39%
        42.2
        HC33HDMXY_n01_T0_G06
        74.0%
        40%
        40.2
        HC33HDMXY_n01_T0_G10
        79.4%
        40%
        61.1
        HC33HDMXY_n01_T0_H03
        78.8%
        40%
        58.5
        HC33HDMXY_n01_T0_H09
        71.6%
        40%
        34.7
        HC33HDMXY_n01_T18_A04
        78.2%
        40%
        54.7
        HC33HDMXY_n01_T18_A06
        73.4%
        39%
        39.1
        HC33HDMXY_n01_T18_B03
        77.6%
        40%
        51.2
        HC33HDMXY_n01_T18_B09
        78.7%
        40%
        60.9
        HC33HDMXY_n01_T18_C04
        77.9%
        40%
        53.0
        HC33HDMXY_n01_T18_C06
        72.0%
        39%
        34.5
        HC33HDMXY_n01_T18_D03
        79.3%
        39%
        59.1
        HC33HDMXY_n01_T18_D09
        76.5%
        40%
        51.3
        HC33HDMXY_n01_T18_E04
        74.7%
        40%
        45.7
        HC33HDMXY_n01_T18_E06
        70.8%
        39%
        32.9
        HC33HDMXY_n01_T18_F03
        78.2%
        39%
        53.7
        HC33HDMXY_n01_T18_F09
        77.8%
        39%
        56.0
        HC33HDMXY_n01_T18_G04
        74.6%
        40%
        43.8
        HC33HDMXY_n01_T18_G06
        68.5%
        39%
        28.8
        HC33HDMXY_n01_T18_H03
        76.2%
        40%
        47.5
        HC33HDMXY_n01_T18_H09
        72.5%
        39%
        35.0
        HC33HDMXY_n01_T4_A04
        77.6%
        40%
        50.8
        HC33HDMXY_n01_T4_A06
        79.2%
        40%
        62.6
        HC33HDMXY_n01_T4_A10
        75.8%
        40%
        47.3
        HC33HDMXY_n01_T4_B03
        76.6%
        40%
        48.3
        HC33HDMXY_n01_T4_B09
        80.1%
        40%
        64.5
        HC33HDMXY_n01_T4_C04
        48.3%
        40%
        17.9
        HC33HDMXY_n01_T4_C06
        75.0%
        40%
        43.1
        HC33HDMXY_n01_T4_C10
        77.2%
        40%
        52.2
        HC33HDMXY_n01_T4_D03
        74.0%
        40%
        38.3
        HC33HDMXY_n01_T4_D09
        79.0%
        40%
        59.3
        HC33HDMXY_n01_T4_E04
        77.2%
        40%
        50.0
        HC33HDMXY_n01_T4_E06
        72.0%
        40%
        35.5
        HC33HDMXY_n01_T4_E10
        69.5%
        41%
        29.5
        HC33HDMXY_n01_T4_F03
        72.4%
        41%
        36.3
        HC33HDMXY_n01_T4_F09
        79.0%
        41%
        64.4
        HC33HDMXY_n01_T4_G04
        74.0%
        40%
        39.9
        HC33HDMXY_n01_T4_G06
        79.4%
        40%
        61.5
        HC33HDMXY_n01_T4_G10
        76.9%
        40%
        48.7
        HC33HDMXY_n01_T4_H03
        72.9%
        40%
        39.9
        HC33HDMXY_n01_T4_H09
        75.6%
        40%
        46.4
        HC33HDMXY_n01_T6_A04
        80.0%
        40%
        64.7
        HC33HDMXY_n01_T6_A06
        69.7%
        40%
        33.8
        HC33HDMXY_n01_T6_A10
        72.6%
        39%
        38.0
        HC33HDMXY_n01_T6_B03
        78.1%
        40%
        54.9
        HC33HDMXY_n01_T6_B09
        78.4%
        40%
        56.7
        HC33HDMXY_n01_T6_C04
        79.1%
        39%
        58.9
        HC33HDMXY_n01_T6_C06
        77.0%
        40%
        51.6
        HC33HDMXY_n01_T6_C10
        72.2%
        39%
        36.2
        HC33HDMXY_n01_T6_D03
        82.5%
        40%
        85.9
        HC33HDMXY_n01_T6_D09
        75.5%
        39%
        43.2
        HC33HDMXY_n01_T6_E04
        77.3%
        40%
        52.6
        HC33HDMXY_n01_T6_E06
        72.8%
        40%
        37.8
        HC33HDMXY_n01_T6_E10
        68.7%
        40%
        30.5
        HC33HDMXY_n01_T6_F03
        78.2%
        44%
        37.2
        HC33HDMXY_n01_T6_F09
        76.3%
        39%
        48.4
        HC33HDMXY_n01_T6_G04
        77.0%
        41%
        56.5
        HC33HDMXY_n01_T6_G06
        77.8%
        39%
        53.0
        HC33HDMXY_n01_T6_G10
        73.8%
        40%
        41.2
        HC33HDMXY_n01_T6_H03
        81.3%
        40%
        64.7
        HC33HDMXY_n01_T6_H09
        72.9%
        39%
        36.8
        HC33HDMXY_n01_T8_A04
        76.2%
        39%
        47.7
        HC33HDMXY_n01_T8_A06
        72.3%
        39%
        35.7
        HC33HDMXY_n01_T8_A10
        76.7%
        39%
        54.3
        HC33HDMXY_n01_T8_B03
        76.4%
        40%
        51.9
        HC33HDMXY_n01_T8_B09
        75.9%
        40%
        48.3
        HC33HDMXY_n01_T8_C04
        79.3%
        40%
        63.3
        HC33HDMXY_n01_T8_C06
        77.8%
        40%
        56.3
        HC33HDMXY_n01_T8_C10
        73.9%
        40%
        43.0
        HC33HDMXY_n01_T8_D03
        76.5%
        41%
        49.7
        HC33HDMXY_n01_T8_D09
        76.9%
        40%
        51.1
        HC33HDMXY_n01_T8_E04
        71.2%
        39%
        34.1
        HC33HDMXY_n01_T8_E06
        75.7%
        40%
        48.0
        HC33HDMXY_n01_T8_E10
        75.5%
        43%
        37.5
        HC33HDMXY_n01_T8_F03
        72.5%
        40%
        35.6
        HC33HDMXY_n01_T8_F09
        78.5%
        39%
        58.6
        HC33HDMXY_n01_T8_G04
        73.9%
        40%
        41.3
        HC33HDMXY_n01_T8_G06
        78.9%
        40%
        62.2
        HC33HDMXY_n01_T8_G10
        74.9%
        40%
        45.7
        HC33HDMXY_n01_T8_H03
        74.8%
        40%
        43.1
        HC33HDMXY_n01_T8_H09
        75.5%
        39%
        45.2
        HC33HDMXY_n01_undetermined
        86.5%
        42%
        328.8
        HC33HDMXY_n02_T0_A04
        76.2%
        40%
        52.1
        HC33HDMXY_n02_T0_A06
        74.5%
        40%
        45.5
        HC33HDMXY_n02_T0_A10
        74.0%
        39%
        45.2
        HC33HDMXY_n02_T0_B03
        76.0%
        40%
        51.8
        HC33HDMXY_n02_T0_B09
        76.4%
        40%
        49.9
        HC33HDMXY_n02_T0_C04
        73.6%
        40%
        40.0
        HC33HDMXY_n02_T0_C06
        75.1%
        40%
        43.9
        HC33HDMXY_n02_T0_C10
        76.6%
        39%
        56.6
        HC33HDMXY_n02_T0_D03
        76.6%
        39%
        51.9
        HC33HDMXY_n02_T0_D09
        75.2%
        40%
        45.6
        HC33HDMXY_n02_T0_E04
        73.7%
        39%
        38.8
        HC33HDMXY_n02_T0_E06
        68.8%
        40%
        27.6
        HC33HDMXY_n02_T0_E10
        74.1%
        40%
        41.3
        HC33HDMXY_n02_T0_F03
        72.0%
        40%
        35.2
        HC33HDMXY_n02_T0_F09
        77.4%
        40%
        52.3
        HC33HDMXY_n02_T0_G04
        73.2%
        39%
        42.2
        HC33HDMXY_n02_T0_G06
        73.4%
        40%
        40.2
        HC33HDMXY_n02_T0_G10
        77.7%
        40%
        61.1
        HC33HDMXY_n02_T0_H03
        77.6%
        40%
        58.5
        HC33HDMXY_n02_T0_H09
        71.1%
        40%
        34.7
        HC33HDMXY_n02_T18_A04
        77.2%
        40%
        54.7
        HC33HDMXY_n02_T18_A06
        71.9%
        39%
        39.1
        HC33HDMXY_n02_T18_B03
        76.1%
        40%
        51.2
        HC33HDMXY_n02_T18_B09
        77.3%
        40%
        60.9
        HC33HDMXY_n02_T18_C04
        77.3%
        40%
        53.0
        HC33HDMXY_n02_T18_C06
        70.5%
        39%
        34.5
        HC33HDMXY_n02_T18_D03
        77.5%
        40%
        59.1
        HC33HDMXY_n02_T18_D09
        75.6%
        40%
        51.3
        HC33HDMXY_n02_T18_E04
        75.8%
        40%
        45.7
        HC33HDMXY_n02_T18_E06
        69.5%
        39%
        32.9
        HC33HDMXY_n02_T18_F03
        76.4%
        40%
        53.7
        HC33HDMXY_n02_T18_F09
        76.5%
        39%
        56.0
        HC33HDMXY_n02_T18_G04
        73.1%
        40%
        43.8
        HC33HDMXY_n02_T18_G06
        67.1%
        39%
        28.8
        HC33HDMXY_n02_T18_H03
        74.3%
        40%
        47.5
        HC33HDMXY_n02_T18_H09
        70.2%
        39%
        35.0
        HC33HDMXY_n02_T4_A04
        76.6%
        40%
        50.8
        HC33HDMXY_n02_T4_A06
        78.3%
        40%
        62.6
        HC33HDMXY_n02_T4_A10
        74.9%
        40%
        47.3
        HC33HDMXY_n02_T4_B03
        75.3%
        40%
        48.3
        HC33HDMXY_n02_T4_B09
        79.1%
        40%
        64.5
        HC33HDMXY_n02_T4_C04
        49.5%
        40%
        17.9
        HC33HDMXY_n02_T4_C06
        74.9%
        40%
        43.1
        HC33HDMXY_n02_T4_C10
        76.6%
        40%
        52.2
        HC33HDMXY_n02_T4_D03
        73.3%
        40%
        38.3
        HC33HDMXY_n02_T4_D09
        77.7%
        40%
        59.3
        HC33HDMXY_n02_T4_E04
        76.3%
        40%
        50.0
        HC33HDMXY_n02_T4_E06
        71.8%
        40%
        35.5
        HC33HDMXY_n02_T4_E10
        69.3%
        41%
        29.5
        HC33HDMXY_n02_T4_F03
        72.4%
        41%
        36.3
        HC33HDMXY_n02_T4_F09
        78.5%
        41%
        64.4
        HC33HDMXY_n02_T4_G04
        73.0%
        40%
        39.9
        HC33HDMXY_n02_T4_G06
        78.3%
        40%
        61.5
        HC33HDMXY_n02_T4_G10
        75.7%
        40%
        48.7
        HC33HDMXY_n02_T4_H03
        72.9%
        40%
        39.9
        HC33HDMXY_n02_T4_H09
        74.7%
        40%
        46.4
        HC33HDMXY_n02_T6_A04
        79.0%
        40%
        64.7
        HC33HDMXY_n02_T6_A06
        71.3%
        40%
        33.8
        HC33HDMXY_n02_T6_A10
        70.6%
        39%
        38.0
        HC33HDMXY_n02_T6_B03
        77.1%
        40%
        54.9
        HC33HDMXY_n02_T6_B09
        77.7%
        40%
        56.7
        HC33HDMXY_n02_T6_C04
        78.2%
        39%
        58.9
        HC33HDMXY_n02_T6_C06
        76.3%
        40%
        51.6
        HC33HDMXY_n02_T6_C10
        70.7%
        39%
        36.2
        HC33HDMXY_n02_T6_D03
        81.7%
        40%
        85.9
        HC33HDMXY_n02_T6_D09
        74.5%
        39%
        43.2
        HC33HDMXY_n02_T6_E04
        77.1%
        40%
        52.6
        HC33HDMXY_n02_T6_E06
        72.7%
        40%
        37.8
        HC33HDMXY_n02_T6_E10
        68.9%
        40%
        30.5
        HC33HDMXY_n02_T6_F03
        78.7%
        44%
        37.2
        HC33HDMXY_n02_T6_F09
        75.9%
        39%
        48.4
        HC33HDMXY_n02_T6_G04
        77.0%
        41%
        56.5
        HC33HDMXY_n02_T6_G06
        76.4%
        39%
        53.0
        HC33HDMXY_n02_T6_G10
        72.3%
        40%
        41.2
        HC33HDMXY_n02_T6_H03
        80.7%
        40%
        64.7
        HC33HDMXY_n02_T6_H09
        71.3%
        39%
        36.8
        HC33HDMXY_n02_T8_A04
        75.5%
        39%
        47.7
        HC33HDMXY_n02_T8_A06
        71.8%
        39%
        35.7
        HC33HDMXY_n02_T8_A10
        75.2%
        39%
        54.3
        HC33HDMXY_n02_T8_B03
        77.3%
        40%
        51.9
        HC33HDMXY_n02_T8_B09
        75.3%
        40%
        48.3
        HC33HDMXY_n02_T8_C04
        78.5%
        39%
        63.3
        HC33HDMXY_n02_T8_C06
        77.3%
        40%
        56.3
        HC33HDMXY_n02_T8_C10
        73.3%
        39%
        43.0
        HC33HDMXY_n02_T8_D03
        76.7%
        41%
        49.7
        HC33HDMXY_n02_T8_D09
        76.2%
        40%
        51.1
        HC33HDMXY_n02_T8_E04
        71.4%
        39%
        34.1
        HC33HDMXY_n02_T8_E06
        75.6%
        40%
        48.0
        HC33HDMXY_n02_T8_E10
        75.6%
        43%
        37.5
        HC33HDMXY_n02_T8_F03
        72.5%
        40%
        35.6
        HC33HDMXY_n02_T8_F09
        77.4%
        39%
        58.6
        HC33HDMXY_n02_T8_G04
        73.1%
        40%
        41.3
        HC33HDMXY_n02_T8_G06
        78.2%
        40%
        62.2
        HC33HDMXY_n02_T8_G10
        73.6%
        40%
        45.7
        HC33HDMXY_n02_T8_H03
        73.7%
        40%
        43.1
        HC33HDMXY_n02_T8_H09
        73.8%
        39%
        45.2
        HC33HDMXY_n02_undetermined
        84.3%
        42%
        328.8

        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 97/97 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        328834058
        6.8
        T0_B03
        51822969
        1.1
        T0_D03
        51872325
        1.1
        T0_F03
        35181831
        0.7
        T0_H03
        58499449
        1.2
        T0_A04
        52064492
        1.1
        T0_C04
        40016637
        0.8
        T0_E04
        38760512
        0.8
        T0_G04
        42183835
        0.9
        T0_A06
        45511261
        0.9
        T0_C06
        43906350
        0.9
        T0_E06
        27574984
        0.6
        T0_G06
        40169777
        0.8
        T0_B09
        49890851
        1.0
        T0_D09
        45615905
        0.9
        T0_F09
        52285143
        1.1
        T0_H09
        34689859
        0.7
        T0_A10
        45248572
        0.9
        T0_C10
        56592425
        1.2
        T0_E10
        41258583
        0.8
        T0_G10
        61101235
        1.3
        T18_B03
        51175023
        1.1
        T18_D03
        59107638
        1.2
        T18_F03
        53728138
        1.1
        T18_H03
        47501551
        1.0
        T4_B03
        48317169
        1.0
        T4_D03
        38263171
        0.8
        T4_F03
        36330311
        0.7
        T4_H03
        39877318
        0.8
        T4_A04
        50775449
        1.0
        T4_C04
        17919282
        0.4
        T4_E04
        49979706
        1.0
        T4_G04
        39896157
        0.8
        T4_A06
        62578235
        1.3
        T4_C06
        43077554
        0.9
        T4_E06
        35546636
        0.7
        T4_G06
        61497757
        1.3
        T4_B09
        64463456
        1.3
        T4_D09
        59311280
        1.2
        T4_F09
        64384116
        1.3
        T4_H09
        46407299
        1.0
        T4_A10
        47348669
        1.0
        T4_C10
        52167819
        1.1
        T4_E10
        29522393
        0.6
        T4_G10
        48720583
        1.0
        T18_A04
        54697622
        1.1
        T18_C04
        53039771
        1.1
        T18_E04
        45650536
        0.9
        T18_G04
        43839992
        0.9
        T6_B03
        54945583
        1.1
        T6_D03
        85914845
        1.8
        T6_F03
        37230573
        0.8
        T6_H03
        64724718
        1.3
        T6_A04
        64722739
        1.3
        T6_C04
        58927146
        1.2
        T6_E04
        52649032
        1.1
        T6_G04
        56474815
        1.2
        T6_A06
        33830306
        0.7
        T6_C06
        51594356
        1.1
        T6_E06
        37848472
        0.8
        T6_G06
        52983226
        1.1
        T6_B09
        56686426
        1.2
        T6_D09
        43196617
        0.9
        T6_F09
        48424595
        1.0
        T6_H09
        36843466
        0.8
        T6_A10
        37956161
        0.8
        T6_C10
        36211659
        0.7
        T6_E10
        30463036
        0.6
        T6_G10
        41193883
        0.8
        T18_A06
        39090795
        0.8
        T18_C06
        34491210
        0.7
        T18_E06
        32920481
        0.7
        T18_G06
        28837994
        0.6
        T8_B03
        51923507
        1.1
        T8_D03
        49678393
        1.0
        T8_F03
        35569177
        0.7
        T8_H03
        43126630
        0.9
        T8_A04
        47671596
        1.0
        T8_C04
        63307819
        1.3
        T8_E04
        34116759
        0.7
        T8_G04
        41285639
        0.8
        T8_A06
        35736380
        0.7
        T8_C06
        56326243
        1.2
        T8_E06
        47989602
        1.0
        T8_G06
        62167964
        1.3
        T8_B09
        48327630
        1.0
        T8_D09
        51057004
        1.1
        T8_F09
        58556650
        1.2
        T8_H09
        45159790
        0.9
        T8_A10
        54330938
        1.1
        T8_C10
        42987804
        0.9
        T8_E10
        37475490
        0.8
        T8_G10
        45734714
        0.9
        T18_B09
        60898772
        1.3
        T18_D09
        51279220
        1.1
        T18_F09
        55993535
        1.2
        T18_H09
        35043874
        0.7

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        2.0
        5761400832
        4862112948
        6.8
        3.2

        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.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGGGGAGATCTCG
        157134155.0
        47.8
        GGGGGGGGTATGCAGT
        4029991.0
        1.2
        GGGGGGGGCTCCTTAC
        3913271.0
        1.2
        GGGGGGGGTACTCCTT
        3834232.0
        1.2
        GGGGGGGGAGGCTTAG
        3566544.0
        1.1
        GGGGGGGGAGAGGATA
        3237600.0
        1.0
        GGGGGGGGATAGAGAG
        2731904.0
        0.8
        GGGGGGGGTCTACTCT
        2632965.0
        0.8
        GGGGGGGGTCTTACGC
        2537821.0
        0.8
        CTCTCTACGGGGGGGG
        1188698.0
        0.4
        AAGAGGCAGGGGGGGG
        1105878.0
        0.3
        GGACTCCTGGGGGGGG
        1055738.0
        0.3
        AGGCAGAAGGGGGGGG
        1031261.0
        0.3
        CGAGGCTGGGGGGGGG
        949290.0
        0.3
        TAAGGCGAGGGGGGGG
        947824.0
        0.3
        GTAGAGGAGGGGGGGG
        926670.0
        0.3
        CAGAGAGGGGGGGGGG
        902295.0
        0.3
        TAGGCATGGGGGGGGG
        880777.0
        0.3
        CGTACTAGGGGGGGGG
        841278.0
        0.3
        TCCTGAGCGGGGGGGG
        823500.0
        0.2

        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.

        194 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.

        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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