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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 2022-04-10, 22:03 based on data in: /scratch/gencore/logs/html/HC33GDMXY/merged


        General Statistics

        Showing 194/194 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HC33GDMXY_n01_HA_Var
        98.0%
        42%
        53.8
        HC33GDMXY_n01_HA_WT
        98.1%
        42%
        56.2
        HC33GDMXY_n01_NA_Var
        97.9%
        43%
        47.0
        HC33GDMXY_n01_NA_WT
        97.7%
        43%
        40.4
        HC33GDMXY_n01_PB2_Var
        98.0%
        44%
        58.7
        HC33GDMXY_n01_PB2_WT
        97.7%
        44%
        50.7
        HC33GDMXY_n01_T0_H03_R
        68.6%
        39%
        27.3
        HC33GDMXY_n01_T10_A04
        76.0%
        40%
        48.8
        HC33GDMXY_n01_T10_A06
        74.8%
        40%
        44.4
        HC33GDMXY_n01_T10_A10
        73.8%
        39%
        39.3
        HC33GDMXY_n01_T10_B03
        74.5%
        40%
        45.7
        HC33GDMXY_n01_T10_B09
        76.3%
        40%
        49.5
        HC33GDMXY_n01_T10_C04
        77.1%
        40%
        54.2
        HC33GDMXY_n01_T10_C06
        76.1%
        40%
        50.7
        HC33GDMXY_n01_T10_C10
        75.2%
        40%
        46.0
        HC33GDMXY_n01_T10_D03
        75.1%
        40%
        44.2
        HC33GDMXY_n01_T10_D09
        77.3%
        40%
        54.9
        HC33GDMXY_n01_T10_E04
        75.0%
        41%
        45.8
        HC33GDMXY_n01_T10_E06
        74.0%
        40%
        41.8
        HC33GDMXY_n01_T10_E10
        73.5%
        40%
        40.0
        HC33GDMXY_n01_T10_F03
        70.3%
        40%
        24.9
        HC33GDMXY_n01_T10_F09
        69.5%
        40%
        30.2
        HC33GDMXY_n01_T10_G04
        73.1%
        39%
        39.4
        HC33GDMXY_n01_T10_G06
        73.9%
        41%
        42.1
        HC33GDMXY_n01_T10_G10
        74.6%
        40%
        42.7
        HC33GDMXY_n01_T10_H03
        72.1%
        40%
        36.6
        HC33GDMXY_n01_T10_H09
        69.2%
        40%
        30.0
        HC33GDMXY_n01_T12_A04
        73.7%
        40%
        39.8
        HC33GDMXY_n01_T12_A06
        76.3%
        40%
        48.6
        HC33GDMXY_n01_T12_A10
        73.6%
        40%
        42.1
        HC33GDMXY_n01_T12_B03
        75.6%
        40%
        49.2
        HC33GDMXY_n01_T12_B09
        77.9%
        41%
        50.4
        HC33GDMXY_n01_T12_C04
        45.6%
        40%
        17.3
        HC33GDMXY_n01_T12_C06
        77.9%
        40%
        60.3
        HC33GDMXY_n01_T12_C10
        75.9%
        40%
        51.0
        HC33GDMXY_n01_T12_D03
        78.1%
        39%
        58.0
        HC33GDMXY_n01_T12_D09
        76.9%
        40%
        47.2
        HC33GDMXY_n01_T12_E04
        75.8%
        40%
        47.4
        HC33GDMXY_n01_T12_E06
        74.3%
        40%
        43.1
        HC33GDMXY_n01_T12_E10
        75.4%
        40%
        48.4
        HC33GDMXY_n01_T12_F03
        71.2%
        39%
        33.5
        HC33GDMXY_n01_T12_F09
        75.8%
        40%
        46.8
        HC33GDMXY_n01_T12_G04
        73.7%
        40%
        41.1
        HC33GDMXY_n01_T12_G06
        76.2%
        40%
        51.0
        HC33GDMXY_n01_T12_G10
        75.2%
        39%
        44.7
        HC33GDMXY_n01_T12_H03
        71.8%
        40%
        40.5
        HC33GDMXY_n01_T12_H09
        75.6%
        40%
        47.1
        HC33GDMXY_n01_T14_A04
        75.9%
        40%
        47.1
        HC33GDMXY_n01_T14_A06
        73.1%
        40%
        45.6
        HC33GDMXY_n01_T14_A10
        73.8%
        40%
        44.3
        HC33GDMXY_n01_T14_B03
        77.0%
        40%
        52.3
        HC33GDMXY_n01_T14_B09
        75.7%
        40%
        46.8
        HC33GDMXY_n01_T14_C04
        76.1%
        39%
        50.0
        HC33GDMXY_n01_T14_C06
        76.0%
        40%
        50.0
        HC33GDMXY_n01_T14_C10
        72.5%
        39%
        39.3
        HC33GDMXY_n01_T14_D03
        77.9%
        40%
        57.3
        HC33GDMXY_n01_T14_D09
        76.0%
        39%
        48.1
        HC33GDMXY_n01_T14_E04
        76.2%
        40%
        49.6
        HC33GDMXY_n01_T14_E06
        72.9%
        40%
        40.4
        HC33GDMXY_n01_T14_E10
        68.8%
        40%
        30.3
        HC33GDMXY_n01_T14_F03
        75.1%
        40%
        47.0
        HC33GDMXY_n01_T14_F09
        75.1%
        40%
        48.8
        HC33GDMXY_n01_T14_G04
        74.2%
        41%
        45.4
        HC33GDMXY_n01_T14_G06
        73.9%
        40%
        40.6
        HC33GDMXY_n01_T14_G10
        74.7%
        40%
        45.4
        HC33GDMXY_n01_T14_H03
        76.9%
        39%
        52.8
        HC33GDMXY_n01_T14_H09
        74.2%
        41%
        46.7
        HC33GDMXY_n01_T16_A04
        74.3%
        41%
        52.4
        HC33GDMXY_n01_T16_A06
        71.5%
        41%
        44.9
        HC33GDMXY_n01_T16_A10
        72.9%
        44%
        47.6
        HC33GDMXY_n01_T16_B03
        71.9%
        42%
        37.9
        HC33GDMXY_n01_T16_B09
        71.4%
        41%
        36.7
        HC33GDMXY_n01_T16_C04
        78.4%
        42%
        62.0
        HC33GDMXY_n01_T16_C06
        76.3%
        41%
        53.5
        HC33GDMXY_n01_T16_C10
        65.5%
        43%
        31.0
        HC33GDMXY_n01_T16_D03
        74.0%
        41%
        46.8
        HC33GDMXY_n01_T16_D09
        71.9%
        42%
        39.7
        HC33GDMXY_n01_T16_E04
        78.4%
        43%
        60.3
        HC33GDMXY_n01_T16_E06
        70.5%
        41%
        38.6
        HC33GDMXY_n01_T16_E10
        72.9%
        41%
        45.8
        HC33GDMXY_n01_T16_F03
        73.8%
        41%
        48.3
        HC33GDMXY_n01_T16_F09
        77.3%
        42%
        59.0
        HC33GDMXY_n01_T16_G04
        75.7%
        42%
        49.4
        HC33GDMXY_n01_T16_G06
        74.3%
        41%
        52.5
        HC33GDMXY_n01_T16_H03
        79.7%
        43%
        68.4
        HC33GDMXY_n01_T16_H09
        77.0%
        45%
        62.5
        HC33GDMXY_n01_T18_A10
        76.4%
        40%
        50.9
        HC33GDMXY_n01_T18_C10
        76.0%
        39%
        45.5
        HC33GDMXY_n01_T18_D3_R
        71.4%
        39%
        34.3
        HC33GDMXY_n01_T18_E10
        74.9%
        39%
        42.1
        HC33GDMXY_n01_T18_H09_R
        71.3%
        39%
        33.7
        HC33GDMXY_n01_T4_C6_R
        72.5%
        39%
        39.1
        HC33GDMXY_n01_T6_A10_R
        75.6%
        39%
        48.0
        HC33GDMXY_n01_T6_F03_R
        80.8%
        43%
        60.5
        HC33GDMXY_n01_T6_G04_R
        77.6%
        40%
        56.7
        HC33GDMXY_n01_T8_E10_R
        79.2%
        43%
        58.5
        HC33GDMXY_n01_undetermined
        86.5%
        42%
        365.2
        HC33GDMXY_n02_HA_Var
        97.5%
        42%
        53.8
        HC33GDMXY_n02_HA_WT
        97.3%
        42%
        56.2
        HC33GDMXY_n02_NA_Var
        97.1%
        43%
        47.0
        HC33GDMXY_n02_NA_WT
        97.0%
        43%
        40.4
        HC33GDMXY_n02_PB2_Var
        97.1%
        44%
        58.7
        HC33GDMXY_n02_PB2_WT
        96.7%
        44%
        50.7
        HC33GDMXY_n02_T0_H03_R
        66.5%
        39%
        27.3
        HC33GDMXY_n02_T10_A04
        74.3%
        40%
        48.8
        HC33GDMXY_n02_T10_A06
        74.1%
        40%
        44.4
        HC33GDMXY_n02_T10_A10
        71.2%
        39%
        39.3
        HC33GDMXY_n02_T10_B03
        73.4%
        40%
        45.7
        HC33GDMXY_n02_T10_B09
        75.6%
        40%
        49.5
        HC33GDMXY_n02_T10_C04
        76.3%
        40%
        54.2
        HC33GDMXY_n02_T10_C06
        75.9%
        40%
        50.7
        HC33GDMXY_n02_T10_C10
        73.3%
        40%
        46.0
        HC33GDMXY_n02_T10_D03
        73.9%
        40%
        44.2
        HC33GDMXY_n02_T10_D09
        76.5%
        40%
        54.9
        HC33GDMXY_n02_T10_E04
        76.9%
        41%
        45.8
        HC33GDMXY_n02_T10_E06
        73.8%
        40%
        41.8
        HC33GDMXY_n02_T10_E10
        72.7%
        40%
        40.0
        HC33GDMXY_n02_T10_F03
        69.6%
        40%
        24.9
        HC33GDMXY_n02_T10_F09
        69.7%
        40%
        30.2
        HC33GDMXY_n02_T10_G04
        71.5%
        40%
        39.4
        HC33GDMXY_n02_T10_G06
        73.3%
        41%
        42.1
        HC33GDMXY_n02_T10_G10
        72.3%
        40%
        42.7
        HC33GDMXY_n02_T10_H03
        70.6%
        40%
        36.6
        HC33GDMXY_n02_T10_H09
        68.6%
        40%
        30.0
        HC33GDMXY_n02_T12_A04
        72.0%
        39%
        39.8
        HC33GDMXY_n02_T12_A06
        74.9%
        40%
        48.6
        HC33GDMXY_n02_T12_A10
        72.1%
        40%
        42.1
        HC33GDMXY_n02_T12_B03
        73.6%
        40%
        49.2
        HC33GDMXY_n02_T12_B09
        76.6%
        41%
        50.4
        HC33GDMXY_n02_T12_C04
        42.8%
        40%
        17.3
        HC33GDMXY_n02_T12_C06
        77.3%
        40%
        60.3
        HC33GDMXY_n02_T12_C10
        75.0%
        40%
        51.0
        HC33GDMXY_n02_T12_D03
        76.8%
        39%
        58.0
        HC33GDMXY_n02_T12_D09
        74.9%
        40%
        47.2
        HC33GDMXY_n02_T12_E04
        74.6%
        40%
        47.4
        HC33GDMXY_n02_T12_E06
        73.8%
        40%
        43.1
        HC33GDMXY_n02_T12_E10
        74.7%
        40%
        48.4
        HC33GDMXY_n02_T12_F03
        70.7%
        39%
        33.5
        HC33GDMXY_n02_T12_F09
        74.2%
        40%
        46.8
        HC33GDMXY_n02_T12_G04
        71.8%
        40%
        41.1
        HC33GDMXY_n02_T12_G06
        74.6%
        40%
        51.0
        HC33GDMXY_n02_T12_G10
        73.0%
        39%
        44.7
        HC33GDMXY_n02_T12_H03
        70.4%
        40%
        40.5
        HC33GDMXY_n02_T12_H09
        73.9%
        40%
        47.1
        HC33GDMXY_n02_T14_A04
        74.5%
        40%
        47.1
        HC33GDMXY_n02_T14_A06
        74.7%
        40%
        45.6
        HC33GDMXY_n02_T14_A10
        71.5%
        40%
        44.3
        HC33GDMXY_n02_T14_B03
        75.0%
        40%
        52.3
        HC33GDMXY_n02_T14_B09
        74.5%
        40%
        46.8
        HC33GDMXY_n02_T14_C04
        75.2%
        39%
        50.0
        HC33GDMXY_n02_T14_C06
        75.2%
        40%
        50.0
        HC33GDMXY_n02_T14_C10
        71.0%
        39%
        39.3
        HC33GDMXY_n02_T14_D03
        76.8%
        40%
        57.3
        HC33GDMXY_n02_T14_D09
        74.7%
        39%
        48.1
        HC33GDMXY_n02_T14_E04
        75.7%
        40%
        49.6
        HC33GDMXY_n02_T14_E06
        73.0%
        40%
        40.4
        HC33GDMXY_n02_T14_E10
        68.9%
        40%
        30.3
        HC33GDMXY_n02_T14_F03
        75.1%
        40%
        47.0
        HC33GDMXY_n02_T14_F09
        74.9%
        40%
        48.8
        HC33GDMXY_n02_T14_G04
        73.4%
        41%
        45.4
        HC33GDMXY_n02_T14_G06
        72.0%
        40%
        40.6
        HC33GDMXY_n02_T14_G10
        72.6%
        40%
        45.4
        HC33GDMXY_n02_T14_H03
        75.5%
        40%
        52.8
        HC33GDMXY_n02_T14_H09
        73.2%
        40%
        46.7
        HC33GDMXY_n02_T16_A04
        73.4%
        41%
        52.4
        HC33GDMXY_n02_T16_A06
        71.4%
        41%
        44.9
        HC33GDMXY_n02_T16_A10
        72.0%
        43%
        47.6
        HC33GDMXY_n02_T16_B03
        72.3%
        42%
        37.9
        HC33GDMXY_n02_T16_B09
        69.4%
        41%
        36.7
        HC33GDMXY_n02_T16_C04
        77.9%
        42%
        62.0
        HC33GDMXY_n02_T16_C06
        75.3%
        41%
        53.5
        HC33GDMXY_n02_T16_C10
        64.2%
        43%
        31.0
        HC33GDMXY_n02_T16_D03
        73.9%
        41%
        46.8
        HC33GDMXY_n02_T16_D09
        70.4%
        42%
        39.7
        HC33GDMXY_n02_T16_E04
        78.7%
        42%
        60.3
        HC33GDMXY_n02_T16_E06
        70.1%
        41%
        38.6
        HC33GDMXY_n02_T16_E10
        72.1%
        41%
        45.8
        HC33GDMXY_n02_T16_F03
        73.6%
        41%
        48.3
        HC33GDMXY_n02_T16_F09
        76.9%
        42%
        59.0
        HC33GDMXY_n02_T16_G04
        74.9%
        42%
        49.4
        HC33GDMXY_n02_T16_G06
        73.2%
        41%
        52.5
        HC33GDMXY_n02_T16_H03
        78.9%
        42%
        68.4
        HC33GDMXY_n02_T16_H09
        76.0%
        45%
        62.5
        HC33GDMXY_n02_T18_A10
        74.2%
        40%
        50.9
        HC33GDMXY_n02_T18_C10
        73.5%
        39%
        45.5
        HC33GDMXY_n02_T18_D3_R
        69.7%
        39%
        34.3
        HC33GDMXY_n02_T18_E10
        72.6%
        40%
        42.1
        HC33GDMXY_n02_T18_H09_R
        69.3%
        39%
        33.7
        HC33GDMXY_n02_T4_C6_R
        70.6%
        40%
        39.1
        HC33GDMXY_n02_T6_A10_R
        73.8%
        39%
        48.0
        HC33GDMXY_n02_T6_F03_R
        81.5%
        43%
        60.5
        HC33GDMXY_n02_T6_G04_R
        76.2%
        40%
        56.7
        HC33GDMXY_n02_T8_E10_R
        78.4%
        43%
        58.5
        HC33GDMXY_n02_undetermined
        84.0%
        42%
        365.2

        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
        365219778
        7.6
        T10_B03
        45690214
        1.0
        T10_D03
        44204601
        0.9
        T10_F03
        24930047
        0.5
        T10_H03
        36573780
        0.8
        T10_A04
        48818667
        1.0
        T10_C04
        54246582
        1.1
        T10_E04
        45822121
        1.0
        T10_G04
        39425015
        0.8
        T10_A06
        44381874
        0.9
        T10_C06
        50684612
        1.1
        T10_E06
        41829217
        0.9
        T10_G06
        42115017
        0.9
        T10_B09
        49455603
        1.0
        T10_D09
        54944080
        1.1
        T10_F09
        30163182
        0.6
        T10_H09
        29987369
        0.6
        T10_A10
        39295557
        0.8
        T10_C10
        45986618
        1.0
        T10_E10
        39983918
        0.8
        T10_G10
        42681744
        0.9
        T18_A10
        50933437
        1.1
        T18_C10
        45495020
        0.9
        T18_E10
        42051560
        0.9
        NA_Var
        47049243
        1.0
        T12_B03
        49195584
        1.0
        T12_D03
        57988274
        1.2
        T12_F03
        33503608
        0.7
        T12_H03
        40547604
        0.8
        T12_A04
        39804522
        0.8
        T12_C04
        17265629
        0.4
        T12_E04
        47382675
        1.0
        T12_G04
        41141945
        0.9
        T12_A06
        48564920
        1.0
        T12_C06
        60307945
        1.3
        T12_E06
        43148536
        0.9
        T12_G06
        50953029
        1.1
        T12_B09
        50403113
        1.1
        T12_D09
        47200378
        1.0
        T12_F09
        46765949
        1.0
        T12_H09
        47058768
        1.0
        T12_A10
        42137049
        0.9
        T12_C10
        50991650
        1.1
        T12_E10
        48388124
        1.0
        T12_G10
        44663399
        0.9
        T6_G04_R
        56653575
        1.2
        T6_A10_R
        48006891
        1.0
        T6_F03_R
        60546046
        1.3
        T8_E10_R
        58514770
        1.2
        T14_B03
        52307195
        1.1
        T14_D03
        57343177
        1.2
        T14_F03
        46963586
        1.0
        T14_H03
        52842287
        1.1
        T14_A04
        47109490
        1.0
        T14_C04
        50047762
        1.0
        T14_E04
        49582449
        1.0
        T14_G04
        45358661
        0.9
        T14_A06
        45629245
        1.0
        T14_C06
        50006538
        1.0
        T14_E06
        40388014
        0.8
        T14_G06
        40555531
        0.8
        T14_B09
        46802563
        1.0
        T14_D09
        48147028
        1.0
        T14_F09
        48806479
        1.0
        T14_H09
        46685932
        1.0
        T14_A10
        44282928
        0.9
        T14_C10
        39266511
        0.8
        T14_E10
        30269622
        0.6
        T14_G10
        45402412
        0.9
        T18_H09_R
        33663159
        0.7
        T0_H03_R
        27268232
        0.6
        T18_D3_R
        34346578
        0.7
        T4_C6_R
        39114390
        0.8
        T16_B03
        37900263
        0.8
        T16_D03
        46751457
        1.0
        T16_F03
        48340114
        1.0
        T16_H03
        68383626
        1.4
        T16_A04
        52388063
        1.1
        T16_C04
        61950688
        1.3
        T16_E04
        60330723
        1.3
        T16_G04
        49382701
        1.0
        T16_A06
        44874513
        0.9
        T16_C06
        53499648
        1.1
        T16_E06
        38609827
        0.8
        T16_G06
        52513371
        1.1
        T16_B09
        36661608
        0.8
        T16_D09
        39677097
        0.8
        T16_F09
        59023015
        1.2
        T16_H09
        62486046
        1.3
        T16_A10
        47618248
        1.0
        T16_C10
        31049434
        0.6
        T16_E10
        45766929
        1.0
        PB2_WT
        50749094
        1.1
        PB2_Var
        58699501
        1.2
        HA_WT
        56162139
        1.2
        HA_Var
        53769488
        1.1
        NA_WT
        40423880
        0.8

        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
        4792304081
        7.6
        4.0

        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
        189388994.0
        51.9
        GGGGGGGGAGGCTTAG
        4057991.0
        1.1
        GGGGGGGGTATGCAGT
        3966278.0
        1.1
        GGGGGGGGTACTCCTT
        3933851.0
        1.1
        GGGGGGGGAGAGGATA
        3694206.0
        1.0
        GGGGGGGGTCTTACGC
        3465918.0
        0.9
        GGGGGGGGCTCCTTAC
        3394867.0
        0.9
        GGGGGGGGATAGAGAG
        2958350.0
        0.8
        GGGGGGGGTCTACTCT
        2935841.0
        0.8
        CGAGGCTGGGGGGGGG
        1074466.0
        0.3
        CTCTCTACGGGGGGGG
        1031797.0
        0.3
        TAGGCATGGGGGGGGG
        1014044.0
        0.3
        AAGAGGCAGGGGGGGG
        1009325.0
        0.3
        GGACTCCTGGGGGGGG
        1004639.0
        0.3
        GTAGAGGAGGGGGGGG
        931385.0
        0.3
        AGGCAGAAGGGGGGGG
        912206.0
        0.2
        TCCTGAGCGGGGGGGG
        903272.0
        0.2
        CGTACTAGGGGGGGGG
        865269.0
        0.2
        CAGAGAGGGGGGGGGG
        858235.0
        0.2
        TAAGGCGAGGGGGGGG
        818071.0
        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.

        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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