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        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 1.18

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        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 2026-04-22, 02:46 CDT based on data in: /scratch/g/akwitek/wdemos/GSE98520


        General Statistics

        Showing 266/266 rows and 6/9 columns.
        Sample Name% Alignable, M% AlignedM Aligned% Dups% GCM Seqs
        GSM2598175
        94.6%
        GSM2598175_SRR5504212_1
        63.8%
        46%
        4.0
        GSM2598175_SRR5504212_2
        63.0%
        46%
        4.0
        GSM2598175_SRR5504213_1
        60.5%
        46%
        4.0
        GSM2598175_SRR5504213_2
        59.9%
        46%
        4.0
        GSM2598175_SRR5504214_1
        63.3%
        46%
        4.0
        GSM2598175_SRR5504214_2
        62.1%
        46%
        4.0
        GSM2598175_SRR5504215_1
        63.9%
        46%
        4.0
        GSM2598175_SRR5504215_2
        64.1%
        46%
        4.0
        GSM2598175_SRR5504216_1
        63.9%
        46%
        4.0
        GSM2598175_SRR5504216_2
        64.2%
        46%
        4.0
        GSM2598175_SRR5504217_1
        64.1%
        46%
        4.0
        GSM2598175_SRR5504217_2
        63.8%
        46%
        4.0
        GSM2598175_SRR5504218_1
        57.6%
        46%
        1.6
        GSM2598175_SRR5504218_2
        57.1%
        46%
        1.6
        GSM2598175_STAR
        92.6%
        23.7
        GSM2598176
        93.7%
        GSM2598176_SRR5504219_1
        60.4%
        47%
        4.0
        GSM2598176_SRR5504219_2
        60.1%
        47%
        4.0
        GSM2598176_SRR5504220_1
        60.2%
        47%
        4.0
        GSM2598176_SRR5504220_2
        60.5%
        47%
        4.0
        GSM2598176_SRR5504221_1
        60.4%
        47%
        4.0
        GSM2598176_SRR5504221_2
        60.7%
        47%
        4.0
        GSM2598176_SRR5504222_1
        60.6%
        47%
        4.0
        GSM2598176_SRR5504222_2
        61.3%
        47%
        4.0
        GSM2598176_SRR5504223_1
        60.7%
        47%
        4.0
        GSM2598176_SRR5504223_2
        61.4%
        47%
        4.0
        GSM2598176_SRR5504224_1
        61.1%
        47%
        4.0
        GSM2598176_SRR5504224_2
        61.7%
        47%
        4.0
        GSM2598176_SRR5504225_1
        61.0%
        47%
        4.0
        GSM2598176_SRR5504225_2
        60.4%
        47%
        4.0
        GSM2598176_SRR5504226_1
        45.6%
        47%
        0.2
        GSM2598176_SRR5504226_2
        46.8%
        47%
        0.2
        GSM2598176_STAR
        91.9%
        25.9
        GSM2598177
        94.3%
        GSM2598177_SRR5504227_1
        62.6%
        46%
        4.0
        GSM2598177_SRR5504227_2
        61.6%
        46%
        4.0
        GSM2598177_SRR5504228_1
        62.9%
        46%
        4.0
        GSM2598177_SRR5504228_2
        61.5%
        46%
        4.0
        GSM2598177_SRR5504229_1
        62.7%
        46%
        4.0
        GSM2598177_SRR5504229_2
        62.1%
        46%
        4.0
        GSM2598177_SRR5504230_1
        62.9%
        46%
        4.0
        GSM2598177_SRR5504230_2
        62.6%
        46%
        4.0
        GSM2598177_SRR5504231_1
        62.6%
        46%
        4.0
        GSM2598177_SRR5504231_2
        62.2%
        46%
        4.0
        GSM2598177_SRR5504232_1
        61.3%
        46%
        4.0
        GSM2598177_SRR5504232_2
        60.8%
        46%
        4.0
        GSM2598177_SRR5504233_1
        42.3%
        46%
        0.3
        GSM2598177_SRR5504233_2
        43.8%
        46%
        0.3
        GSM2598177_STAR
        92.2%
        22.4
        GSM2598178
        93.3%
        GSM2598178_SRR5504234_1
        63.4%
        46%
        4.0
        GSM2598178_SRR5504234_2
        62.7%
        46%
        4.0
        GSM2598178_SRR5504235_1
        63.2%
        46%
        4.0
        GSM2598178_SRR5504235_2
        62.8%
        46%
        4.0
        GSM2598178_SRR5504236_1
        63.6%
        46%
        4.0
        GSM2598178_SRR5504236_2
        63.1%
        46%
        4.0
        GSM2598178_SRR5504237_1
        63.5%
        46%
        4.0
        GSM2598178_SRR5504237_2
        63.5%
        46%
        4.0
        GSM2598178_SRR5504238_1
        63.6%
        46%
        4.0
        GSM2598178_SRR5504238_2
        63.7%
        46%
        4.0
        GSM2598178_SRR5504239_1
        63.9%
        46%
        4.0
        GSM2598178_SRR5504239_2
        64.0%
        46%
        4.0
        GSM2598178_SRR5504240_1
        64.0%
        46%
        4.0
        GSM2598178_SRR5504240_2
        62.8%
        46%
        4.0
        GSM2598178_SRR5504241_1
        43.3%
        46%
        0.3
        GSM2598178_SRR5504241_2
        45.1%
        46%
        0.3
        GSM2598178_STAR
        91.4%
        25.8
        GSM2598179
        92.1%
        GSM2598179_SRR5504242_1
        56.5%
        46%
        4.0
        GSM2598179_SRR5504242_2
        55.7%
        46%
        4.0
        GSM2598179_SRR5504243_1
        56.3%
        46%
        4.0
        GSM2598179_SRR5504243_2
        55.4%
        46%
        4.0
        GSM2598179_SRR5504244_1
        56.5%
        46%
        4.0
        GSM2598179_SRR5504244_2
        56.1%
        46%
        4.0
        GSM2598179_SRR5504245_1
        56.5%
        46%
        4.0
        GSM2598179_SRR5504245_2
        56.2%
        46%
        4.0
        GSM2598179_SRR5504246_1
        56.8%
        46%
        4.0
        GSM2598179_SRR5504246_2
        56.7%
        46%
        4.0
        GSM2598179_SRR5504247_1
        56.3%
        46%
        4.0
        GSM2598179_SRR5504247_2
        56.3%
        46%
        4.0
        GSM2598179_SRR5504248_1
        56.4%
        46%
        4.0
        GSM2598179_SRR5504248_2
        56.7%
        46%
        4.0
        GSM2598179_STAR
        90.6%
        25.4
        GSM2598180
        93.8%
        GSM2598180_SRR5504249_1
        57.8%
        46%
        4.0
        GSM2598180_SRR5504249_2
        56.5%
        46%
        4.0
        GSM2598180_SRR5504250_1
        55.9%
        46%
        4.0
        GSM2598180_SRR5504250_2
        55.2%
        46%
        4.0
        GSM2598180_SRR5504251_1
        57.5%
        46%
        4.0
        GSM2598180_SRR5504251_2
        56.8%
        46%
        4.0
        GSM2598180_SRR5504252_1
        57.7%
        46%
        4.0
        GSM2598180_SRR5504252_2
        57.0%
        46%
        4.0
        GSM2598180_SRR5504253_1
        58.2%
        46%
        4.0
        GSM2598180_SRR5504253_2
        57.4%
        46%
        4.0
        GSM2598180_SRR5504254_1
        57.7%
        46%
        4.0
        GSM2598180_SRR5504254_2
        56.9%
        46%
        4.0
        GSM2598180_SRR5504255_1
        54.1%
        46%
        2.5
        GSM2598180_SRR5504255_2
        54.4%
        46%
        2.5
        GSM2598180_STAR
        92.0%
        24.4
        GSM2598181
        94.3%
        GSM2598181_SRR5504256_1
        55.4%
        46%
        4.0
        GSM2598181_SRR5504256_2
        55.3%
        46%
        4.0
        GSM2598181_SRR5504257_1
        55.6%
        46%
        4.0
        GSM2598181_SRR5504257_2
        54.8%
        46%
        4.0
        GSM2598181_SRR5504258_1
        55.8%
        46%
        4.0
        GSM2598181_SRR5504258_2
        54.2%
        46%
        4.0
        GSM2598181_SRR5504259_1
        56.0%
        46%
        4.0
        GSM2598181_SRR5504259_2
        55.5%
        46%
        4.0
        GSM2598181_SRR5504260_1
        55.7%
        46%
        4.0
        GSM2598181_SRR5504260_2
        54.4%
        46%
        4.0
        GSM2598181_SRR5504261_1
        54.3%
        46%
        3.4
        GSM2598181_SRR5504261_2
        53.3%
        46%
        3.4
        GSM2598181_STAR
        92.4%
        21.6
        GSM2598182
        94.4%
        GSM2598182_SRR5504262_1
        55.4%
        46%
        4.0
        GSM2598182_SRR5504262_2
        55.3%
        46%
        4.0
        GSM2598182_SRR5504263_1
        55.0%
        46%
        4.0
        GSM2598182_SRR5504263_2
        55.5%
        46%
        4.0
        GSM2598182_SRR5504264_1
        55.5%
        46%
        4.0
        GSM2598182_SRR5504264_2
        55.8%
        46%
        4.0
        GSM2598182_SRR5504265_1
        55.5%
        46%
        4.0
        GSM2598182_SRR5504265_2
        55.9%
        46%
        4.0
        GSM2598182_SRR5504266_1
        56.1%
        46%
        4.0
        GSM2598182_SRR5504266_2
        55.6%
        46%
        4.0
        GSM2598182_SRR5504267_1
        55.1%
        46%
        4.0
        GSM2598182_SRR5504267_2
        54.8%
        46%
        4.0
        GSM2598182_SRR5504268_1
        49.4%
        46%
        4.0
        GSM2598182_SRR5504268_2
        55.9%
        46%
        4.0
        GSM2598182_SRR5504269_1
        53.2%
        46%
        3.1
        GSM2598182_SRR5504269_2
        54.4%
        46%
        3.1
        GSM2598182_STAR
        92.5%
        28.7
        GSM2598183
        94.8%
        GSM2598183_SRR5504270_1
        57.3%
        46%
        4.0
        GSM2598183_SRR5504270_2
        57.2%
        46%
        4.0
        GSM2598183_SRR5504271_1
        54.3%
        46%
        4.0
        GSM2598183_SRR5504271_2
        54.7%
        46%
        4.0
        GSM2598183_SRR5504272_1
        57.0%
        46%
        4.0
        GSM2598183_SRR5504272_2
        57.0%
        46%
        4.0
        GSM2598183_SRR5504273_1
        57.0%
        46%
        4.0
        GSM2598183_SRR5504273_2
        57.4%
        46%
        4.0
        GSM2598183_SRR5504274_1
        57.7%
        46%
        4.0
        GSM2598183_SRR5504274_2
        57.6%
        46%
        4.0
        GSM2598183_SRR5504275_1
        56.2%
        46%
        4.0
        GSM2598183_SRR5504275_2
        57.9%
        46%
        4.0
        GSM2598183_SRR5504276_1
        52.7%
        46%
        2.3
        GSM2598183_SRR5504276_2
        54.1%
        46%
        2.3
        GSM2598183_STAR
        92.9%
        24.4
        GSM2598184
        92.9%
        GSM2598184_SRR5504277_1
        59.1%
        46%
        4.0
        GSM2598184_SRR5504277_2
        58.0%
        46%
        4.0
        GSM2598184_SRR5504278_1
        58.7%
        46%
        4.0
        GSM2598184_SRR5504278_2
        58.1%
        46%
        4.0
        GSM2598184_SRR5504279_1
        59.0%
        46%
        4.0
        GSM2598184_SRR5504279_2
        58.3%
        46%
        4.0
        GSM2598184_SRR5504280_1
        59.0%
        46%
        4.0
        GSM2598184_SRR5504280_2
        58.1%
        46%
        4.0
        GSM2598184_SRR5504281_1
        59.2%
        46%
        4.0
        GSM2598184_SRR5504281_2
        59.0%
        46%
        4.0
        GSM2598184_SRR5504282_1
        58.6%
        46%
        4.0
        GSM2598184_SRR5504282_2
        58.8%
        46%
        4.0
        GSM2598184_SRR5504283_1
        59.0%
        46%
        4.0
        GSM2598184_SRR5504283_2
        58.2%
        46%
        4.0
        GSM2598184_SRR5504284_1
        48.4%
        46%
        0.8
        GSM2598184_SRR5504284_2
        48.3%
        46%
        0.8
        GSM2598184_STAR
        91.1%
        26.2
        GSM2598185
        93.2%
        GSM2598185_SRR5504285_1
        58.2%
        46%
        4.0
        GSM2598185_SRR5504285_2
        57.4%
        46%
        4.0
        GSM2598185_SRR5504286_1
        57.8%
        46%
        4.0
        GSM2598185_SRR5504286_2
        56.9%
        46%
        4.0
        GSM2598185_SRR5504287_1
        58.2%
        46%
        4.0
        GSM2598185_SRR5504287_2
        57.8%
        46%
        4.0
        GSM2598185_SRR5504288_1
        58.1%
        46%
        4.0
        GSM2598185_SRR5504288_2
        57.8%
        46%
        4.0
        GSM2598185_SRR5504289_1
        58.3%
        46%
        4.0
        GSM2598185_SRR5504289_2
        58.2%
        46%
        4.0
        GSM2598185_SRR5504290_1
        57.7%
        46%
        4.0
        GSM2598185_SRR5504290_2
        57.9%
        46%
        4.0
        GSM2598185_SRR5504291_1
        58.1%
        46%
        4.0
        GSM2598185_SRR5504291_2
        56.7%
        46%
        4.0
        GSM2598185_SRR5504292_1
        34.7%
        46%
        0.1
        GSM2598185_SRR5504292_2
        35.2%
        45%
        0.1
        GSM2598185_STAR
        91.6%
        25.7
        GSM2598186
        94.6%
        GSM2598186_SRR5504293_1
        56.2%
        46%
        4.0
        GSM2598186_SRR5504293_2
        55.5%
        46%
        4.0
        GSM2598186_SRR5504294_1
        55.7%
        46%
        4.0
        GSM2598186_SRR5504294_2
        55.7%
        46%
        4.0
        GSM2598186_SRR5504295_1
        56.3%
        46%
        4.0
        GSM2598186_SRR5504295_2
        56.2%
        46%
        4.0
        GSM2598186_SRR5504296_1
        56.2%
        46%
        4.0
        GSM2598186_SRR5504296_2
        56.2%
        46%
        4.0
        GSM2598186_SRR5504297_1
        56.6%
        46%
        4.0
        GSM2598186_SRR5504297_2
        54.9%
        46%
        4.0
        GSM2598186_SRR5504298_1
        55.7%
        46%
        4.0
        GSM2598186_SRR5504298_2
        56.0%
        46%
        4.0
        GSM2598186_SRR5504299_1
        56.3%
        46%
        4.0
        GSM2598186_SRR5504299_2
        55.9%
        46%
        4.0
        GSM2598186_SRR5504300_1
        54.1%
        46%
        2.9
        GSM2598186_SRR5504300_2
        54.4%
        46%
        2.9
        GSM2598186_STAR
        92.5%
        28.6
        GSM2598187
        92.9%
        GSM2598187_SRR5504301_1
        61.4%
        42%
        4.0
        GSM2598187_SRR5504301_2
        56.4%
        42%
        4.0
        GSM2598187_SRR5504302_1
        60.8%
        42%
        4.0
        GSM2598187_SRR5504302_2
        54.9%
        42%
        4.0
        GSM2598187_SRR5504303_1
        60.4%
        42%
        4.0
        GSM2598187_SRR5504303_2
        51.2%
        42%
        4.0
        GSM2598187_SRR5504304_1
        61.0%
        42%
        4.0
        GSM2598187_SRR5504304_2
        59.0%
        42%
        4.0
        GSM2598187_SRR5504305_1
        60.7%
        42%
        4.0
        GSM2598187_SRR5504305_2
        59.7%
        42%
        4.0
        GSM2598187_SRR5504306_1
        61.2%
        42%
        4.0
        GSM2598187_SRR5504306_2
        57.3%
        42%
        4.0
        GSM2598187_SRR5504307_1
        50.0%
        42%
        1.2
        GSM2598187_SRR5504307_2
        52.9%
        42%
        1.2
        GSM2598187_STAR
        92.0%
        23.2
        GSM2598188
        92.8%
        GSM2598188_SRR5504308_1
        57.7%
        43%
        4.0
        GSM2598188_SRR5504308_2
        53.3%
        43%
        4.0
        GSM2598188_SRR5504309_1
        56.7%
        43%
        4.0
        GSM2598188_SRR5504309_2
        47.9%
        43%
        4.0
        GSM2598188_SRR5504310_1
        57.7%
        43%
        4.0
        GSM2598188_SRR5504310_2
        56.3%
        43%
        4.0
        GSM2598188_SRR5504311_1
        57.2%
        43%
        4.0
        GSM2598188_SRR5504311_2
        55.5%
        43%
        4.0
        GSM2598188_SRR5504312_1
        56.9%
        43%
        4.0
        GSM2598188_SRR5504312_2
        54.8%
        43%
        4.0
        GSM2598188_SRR5504313_1
        56.7%
        43%
        4.0
        GSM2598188_SRR5504313_2
        56.3%
        43%
        4.0
        GSM2598188_SRR5504314_1
        51.4%
        43%
        1.8
        GSM2598188_SRR5504314_2
        42.6%
        43%
        1.8
        GSM2598188_STAR
        92.1%
        23.7
        GSM2598189
        92.9%
        GSM2598189_SRR5504315_1
        60.1%
        43%
        4.0
        GSM2598189_SRR5504315_2
        55.6%
        42%
        4.0
        GSM2598189_SRR5504316_1
        59.9%
        43%
        4.0
        GSM2598189_SRR5504316_2
        58.7%
        42%
        4.0
        GSM2598189_SRR5504317_1
        60.0%
        43%
        4.0
        GSM2598189_SRR5504317_2
        58.6%
        42%
        4.0
        GSM2598189_SRR5504318_1
        59.7%
        43%
        4.0
        GSM2598189_SRR5504318_2
        58.6%
        42%
        4.0
        GSM2598189_SRR5504319_1
        59.3%
        43%
        4.0
        GSM2598189_SRR5504319_2
        54.0%
        42%
        4.0
        GSM2598189_SRR5504320_1
        56.4%
        42%
        2.5
        GSM2598189_SRR5504320_2
        55.1%
        42%
        2.5
        GSM2598189_STAR
        91.8%
        20.7
        GSM2598190
        92.1%
        GSM2598190_SRR5504321_1
        63.1%
        41%
        4.0
        GSM2598190_SRR5504321_2
        57.5%
        41%
        4.0
        GSM2598190_SRR5504322_1
        63.0%
        41%
        4.0
        GSM2598190_SRR5504322_2
        60.8%
        41%
        4.0
        GSM2598190_SRR5504323_1
        62.8%
        41%
        4.0
        GSM2598190_SRR5504323_2
        59.6%
        41%
        4.0
        GSM2598190_SRR5504324_1
        62.8%
        41%
        4.0
        GSM2598190_SRR5504324_2
        60.8%
        41%
        4.0
        GSM2598190_SRR5504325_1
        62.9%
        41%
        4.0
        GSM2598190_SRR5504325_2
        60.5%
        41%
        4.0
        GSM2598190_SRR5504326_1
        63.0%
        41%
        4.0
        GSM2598190_SRR5504326_2
        60.5%
        41%
        4.0
        GSM2598190_SRR5504327_1
        60.6%
        41%
        4.0
        GSM2598190_SRR5504327_2
        60.8%
        41%
        4.0
        GSM2598190_SRR5504328_1
        49.8%
        41%
        0.4
        GSM2598190_SRR5504328_2
        46.1%
        41%
        0.4
        GSM2598190_STAR
        91.3%
        25.9

        Rsem

        Rsem RSEM (RNA-Seq by Expectation-Maximization) is a software package forestimating gene and isoform expression levels from RNA-Seq data.DOI: 10.1186/1471-2105-12-323.

        Mapped Reads

        A breakdown of how all reads were aligned for each sample.

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        Multimapping rates

        A frequency histogram showing how many reads were aligned to n reference regions.

        In an ideal world, every sequence reads would align uniquely to a single location in the reference. However, due to factors such as repeititve sequences, short reads and sequencing errors, reads can be align to the reference 0, 1 or more times. This plot shows the frequency of each factor of multimapping. Good samples should have the majority of reads aligning once.

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        STAR

        STAR is an ultrafast universal RNA-seq aligner.DOI: 10.1093/bioinformatics/bts635.

        Alignment Scores

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        FastQ Screen

        Version: 0.15.1

        FastQ Screen allows you to screen a library of sequences in FastQ format against a set of sequence databases so you can see if the composition of the library matches with what you expect.DOI: 10.12688/f1000research.15931.2.

        Mapped Reads

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


        FastQC

        Version: 0.11.9

        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 by sample

        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 overrepresented.

        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 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).


        Top overrepresented sequences

        Top overrepresented sequences across all samples. The table shows 20 most overrepresented sequences across all samples, ranked by the number of samples they occur in.

        Showing 20/20 rows and 3/3 columns.
        Overrepresented sequenceSamplesOccurrences% of all reads
        CAGGGATCGTACTATCTAACTCATCCCTTGACATTGTACTTCATGATACA
        117
        573077
        0.0664%
        GTGGGCTTTTGCTCATGTGTCATTTAGGGTATAGCCTGAGAATAGTGGGA
        117
        686452
        0.0796%
        ATGAAATTGGCTTGAAACCAGTTGTAGGGGGTTCGAATCCTTCCTTTCTT
        117
        595425
        0.0690%
        GTCAGTATCATGCTGCGGCTTCAAATCCGAAATGATGTTTTGATGTGAAG
        115
        614873
        0.0713%
        GGGGGTTCGAATCCTTCCTTTCTTATTTAACTTTTACATAGGAAGGTTCT
        95
        446679
        0.0518%
        CTTCGAATGTGTGGTAAGGTGGGGGGCATCCATGCAGTCATTCTAGGTTA
        74
        322476
        0.0374%
        GTACACTTCTGGGTGGCCGAAGAATCAGAATAGGTGTTGATAAAGAATTG
        73
        348015
        0.0404%
        GTCGGTTTGATGTGACTGTAGCTTGGTTTAGGCGGCCGGGGATTGCGTCG
        57
        251332
        0.0291%
        CCTGGTTTTAGGTCATTGGTTGGGATTATGTAGGAGTCAAAGCATAGGTC
        54
        233169
        0.0270%
        CCTAAATGACACATGAGCAAAAGCCCACTTTGCCATTATATTTGTAGGTG
        50
        203137
        0.0236%
        CTCTTTTCAACTAACCACAAAGATATCGGAACCCTCTACCTATTATTTGG
        47
        201945
        0.0234%
        CCGTAAGTGAGATGAATGAGCCTATAGAGGAGACTGTATTTCATGTGGTA
        42
        168843
        0.0196%
        CTCGTCGTTACTCTGATTATCCAGATGCTTATACCACATGAAATACAGTC
        42
        169390
        0.0196%
        CCAATATCAGACACCTCTCTTTGTATGATCCGTACTAATTACAGCCGTCC
        41
        174111
        0.0202%
        GCCGTAAGTGAGATGAATGAGCCTATAGAGGAGACTGTATTTCATGTGGT
        39
        160505
        0.0186%
        GTTGACTCTTTTCAACTAACCACAAAGATATCGGAACCCTCTACCTATTA
        38
        150614
        0.0175%
        GGCAGATGTAAAGTAGGCTCGGGTGTCTACATCTAGGCCTACTGTGAATA
        37
        156115
        0.0181%
        CGAAGAATCAGAATAGGTGTTGATAAAGAATTGGGTCTCCACCTCCAGCG
        28
        116451
        0.0135%
        GGCGAATACTGCTCCTATAGATAAGACATAGTGGAAGTGAGCTACTACGT
        27
        100548
        0.0117%
        GGGGGATATACTGTTCATCCTGTTCCAGCTCCAGCTTCTACTATGGAGGA
        22
        88121
        0.0102%

        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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        Software Versions

        Software Versions lists versions of software tools extracted from file contents.

        SoftwareVersion
        FastQ Screen0.15.1
        FastQC0.11.9