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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

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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 2020-07-16, 12:57 based on data in:


        General Statistics

        Showing 208/208 rows and 11/14 columns.
        Sample Name% mCpG% mCHG% mCHHM C's% Dups% Aligned% Trimmed% Dups% GCLengthM Seqs
        EPI-103_S27_L005_R1_001
        6.6%
        EPI-103_S27_L005_R1_001_val_1
        26.1%
        2.3%
        1.3%
        183.5
        13.8%
        44.7%
        18.3%
        22%
        78 bp
        20.2
        EPI-103_S27_L005_R2_001
        6.8%
        EPI-103_S27_L005_R2_001_val_2
        27.2%
        22%
        78 bp
        20.2
        EPI-104_S28_L005_R1_001
        8.6%
        EPI-104_S28_L005_R1_001_val_1
        29.5%
        2.4%
        1.4%
        256.7
        14.8%
        45.4%
        19.1%
        22%
        77 bp
        29.6
        EPI-104_S28_L005_R2_001
        8.8%
        EPI-104_S28_L005_R2_001_val_2
        29.1%
        22%
        77 bp
        29.6
        EPI-111_S29_L005_R1_001
        9.3%
        EPI-111_S29_L005_R1_001_val_1
        27.4%
        2.4%
        1.3%
        231.2
        15.0%
        45.6%
        18.9%
        22%
        76 bp
        26.6
        EPI-111_S29_L005_R2_001
        9.4%
        EPI-111_S29_L005_R2_001_val_2
        28.4%
        21%
        76 bp
        26.6
        EPI-113_S30_L005_R1_001
        8.4%
        EPI-113_S30_L005_R1_001_val_1
        27.7%
        2.4%
        1.3%
        224.1
        14.9%
        44.0%
        21.3%
        22%
        77 bp
        25.9
        EPI-113_S30_L005_R2_001
        8.5%
        EPI-113_S30_L005_R2_001_val_2
        31.2%
        22%
        77 bp
        25.9
        EPI-119_S31_L005_R1_001
        7.8%
        EPI-119_S31_L005_R1_001_val_1
        24.9%
        2.3%
        1.3%
        244.0
        15.6%
        42.7%
        22.3%
        22%
        77 bp
        28.5
        EPI-119_S31_L005_R2_001
        7.9%
        EPI-119_S31_L005_R2_001_val_2
        32.4%
        22%
        78 bp
        28.5
        EPI-120_S32_L005_R1_001
        7.8%
        EPI-120_S32_L005_R1_001_val_1
        27.3%
        2.4%
        1.3%
        217.9
        14.3%
        45.8%
        18.8%
        22%
        77 bp
        23.8
        EPI-120_S32_L005_R2_001
        7.9%
        EPI-120_S32_L005_R2_001_val_2
        29.0%
        22%
        78 bp
        23.8
        EPI-127_S33_L005_R1_001
        7.3%
        EPI-127_S33_L005_R1_001_val_1
        27.9%
        2.4%
        1.3%
        196.9
        15.4%
        44.9%
        20.5%
        22%
        78 bp
        22.0
        EPI-127_S33_L005_R2_001
        7.4%
        EPI-127_S33_L005_R2_001_val_2
        30.4%
        22%
        78 bp
        22.0
        EPI-128_S34_L005_R1_001
        8.1%
        EPI-128_S34_L005_R1_001_val_1
        27.9%
        2.4%
        1.3%
        215.1
        15.6%
        44.9%
        21.3%
        22%
        77 bp
        24.2
        EPI-128_S34_L005_R2_001
        8.4%
        EPI-128_S34_L005_R2_001_val_2
        31.5%
        22%
        77 bp
        24.2
        EPI-135_S35_L005_R1_001
        12.2%
        EPI-135_S35_L005_R1_001_val_1
        29.3%
        2.6%
        1.3%
        226.0
        17.0%
        44.3%
        22.4%
        22%
        74 bp
        27.9
        EPI-135_S35_L005_R2_001
        12.3%
        EPI-135_S35_L005_R2_001_val_2
        32.0%
        22%
        74 bp
        27.9
        EPI-136_S36_L005_R1_001
        11.1%
        EPI-136_S36_L005_R1_001_val_1
        26.7%
        2.4%
        1.3%
        222.5
        15.9%
        44.0%
        20.1%
        22%
        74 bp
        27.3
        EPI-136_S36_L005_R2_001
        11.2%
        EPI-136_S36_L005_R2_001_val_2
        29.4%
        22%
        74 bp
        27.3
        EPI-143_S37_L005_R1_001
        8.4%
        EPI-143_S37_L005_R1_001_val_1
        28.2%
        2.4%
        1.3%
        180.0
        15.2%
        44.0%
        18.6%
        22%
        77 bp
        20.9
        EPI-143_S37_L005_R2_001
        8.7%
        EPI-143_S37_L005_R2_001_val_2
        26.8%
        22%
        77 bp
        20.9
        EPI-145_S38_L005_R1_001
        8.4%
        EPI-145_S38_L005_R1_001_val_1
        28.0%
        2.4%
        1.3%
        215.0
        16.4%
        44.2%
        21.5%
        22%
        77 bp
        25.0
        EPI-145_S38_L005_R2_001
        8.6%
        EPI-145_S38_L005_R2_001_val_2
        31.5%
        22%
        77 bp
        25.0
        EPI-151_S2_L002_R1_001
        23.2%
        EPI-151_S2_L002_R1_001_val_1
        30.8%
        2.2%
        1.9%
        154.4
        27.3%
        40.5%
        28.8%
        24%
        76 bp
        24.2
        EPI-151_S2_L002_R2_001
        23.2%
        EPI-151_S2_L002_R2_001_val_2
        37.3%
        23%
        76 bp
        24.2
        EPI-152_S3_L002_R1_001
        9.7%
        EPI-152_S3_L002_R1_001_val_1
        32.0%
        2.5%
        1.9%
        181.7
        28.3%
        39.3%
        29.2%
        23%
        76 bp
        29.5
        EPI-152_S3_L002_R2_001
        9.9%
        EPI-152_S3_L002_R2_001_val_2
        38.1%
        23%
        77 bp
        29.5
        EPI-153_S4_L002_R1_001
        26.1%
        EPI-153_S4_L002_R1_001_val_1
        30.6%
        2.4%
        2.0%
        136.0
        35.8%
        40.1%
        36.6%
        24%
        74 bp
        25.5
        EPI-153_S4_L002_R2_001
        26.1%
        EPI-153_S4_L002_R2_001_val_2
        43.3%
        23%
        74 bp
        25.5
        EPI-154_S5_L002_R1_001
        9.6%
        EPI-154_S5_L002_R1_001_val_1
        30.6%
        2.3%
        2.0%
        142.8
        28.0%
        37.3%
        28.1%
        24%
        76 bp
        25.0
        EPI-154_S5_L002_R2_001
        9.8%
        EPI-154_S5_L002_R2_001_val_2
        35.5%
        23%
        76 bp
        25.0
        EPI-159_S6_L002_R1_001
        11.9%
        EPI-159_S6_L002_R1_001_val_1
        31.4%
        2.6%
        3.0%
        99.1
        27.2%
        41.3%
        29.0%
        23%
        76 bp
        15.2
        EPI-159_S6_L002_R2_001
        12.1%
        EPI-159_S6_L002_R2_001_val_2
        34.6%
        23%
        76 bp
        15.2
        EPI-160_S7_L002_R1_001
        19.8%
        EPI-160_S7_L002_R1_001_val_1
        30.3%
        2.5%
        2.0%
        176.5
        26.6%
        38.4%
        28.7%
        23%
        71 bp
        31.0
        EPI-160_S7_L002_R2_001
        19.9%
        EPI-160_S7_L002_R2_001_val_2
        35.8%
        23%
        71 bp
        31.0
        EPI-161_S8_L002_R1_001
        14.6%
        EPI-161_S8_L002_R1_001_val_1
        28.8%
        2.4%
        2.2%
        124.5
        26.3%
        35.1%
        26.5%
        23%
        72 bp
        24.0
        EPI-161_S8_L002_R2_001
        14.8%
        EPI-161_S8_L002_R2_001_val_2
        31.2%
        23%
        72 bp
        24.0
        EPI-162_S9_L002_R1_001
        16.7%
        EPI-162_S9_L002_R1_001_val_1
        28.5%
        2.4%
        2.3%
        122.9
        22.8%
        36.3%
        23.0%
        24%
        71 bp
        22.2
        EPI-162_S9_L002_R2_001
        16.9%
        EPI-162_S9_L002_R2_001_val_2
        28.3%
        23%
        71 bp
        22.2
        EPI-167_S10_L002_R1_001
        8.5%
        EPI-167_S10_L002_R1_001_val_1
        29.6%
        3.1%
        3.1%
        169.5
        26.9%
        42.6%
        27.4%
        23%
        77 bp
        24.3
        EPI-167_S10_L002_R2_001
        8.7%
        EPI-167_S10_L002_R2_001_val_2
        34.7%
        23%
        77 bp
        24.3
        EPI-168_S11_L002_R1_001
        10.6%
        EPI-168_S11_L002_R1_001_val_1
        30.1%
        2.2%
        2.2%
        139.8
        27.6%
        39.5%
        29.0%
        24%
        76 bp
        22.5
        EPI-168_S11_L002_R2_001
        10.7%
        EPI-168_S11_L002_R2_001_val_2
        37.3%
        23%
        76 bp
        22.5
        EPI-169_S12_L002_R1_001
        15.8%
        EPI-169_S12_L002_R1_001_val_1
        30.4%
        2.4%
        2.5%
        127.2
        23.6%
        39.3%
        25.3%
        24%
        72 bp
        20.8
        EPI-169_S12_L002_R2_001
        15.9%
        EPI-169_S12_L002_R2_001_val_2
        31.7%
        24%
        72 bp
        20.8
        EPI-170_S13_L002_R1_001
        16.1%
        EPI-170_S13_L002_R1_001_val_1
        28.2%
        2.3%
        1.8%
        151.8
        23.8%
        36.6%
        25.4%
        23%
        71 bp
        27.1
        EPI-170_S13_L002_R2_001
        16.3%
        EPI-170_S13_L002_R2_001_val_2
        31.4%
        23%
        71 bp
        27.1
        EPI-175_S14_L003_R1_001
        10.5%
        EPI-175_S14_L003_R1_001_val_1
        29.1%
        2.3%
        2.0%
        169.2
        22.4%
        41.4%
        24.4%
        23%
        76 bp
        24.3
        EPI-175_S14_L003_R2_001
        10.7%
        EPI-175_S14_L003_R2_001_val_2
        33.3%
        23%
        76 bp
        24.3
        EPI-176_S15_L003_R1_001
        9.5%
        EPI-176_S15_L003_R1_001_val_1
        28.2%
        2.1%
        1.7%
        212.7
        26.4%
        37.8%
        28.7%
        23%
        76 bp
        35.9
        EPI-176_S15_L003_R2_001
        9.7%
        EPI-176_S15_L003_R2_001_val_2
        36.3%
        23%
        76 bp
        35.9
        EPI-181_S16_L003_R1_001
        9.0%
        EPI-181_S16_L003_R1_001_val_1
        31.0%
        2.4%
        1.9%
        182.1
        23.9%
        39.7%
        27.0%
        23%
        77 bp
        26.8
        EPI-181_S16_L003_R2_001
        9.2%
        EPI-181_S16_L003_R2_001_val_2
        36.5%
        23%
        77 bp
        26.8
        EPI-182_S17_L003_R1_001
        10.2%
        EPI-182_S17_L003_R1_001_val_1
        29.0%
        2.4%
        1.8%
        222.5
        21.9%
        41.7%
        24.5%
        23%
        76 bp
        31.1
        EPI-182_S17_L003_R2_001
        10.3%
        EPI-182_S17_L003_R2_001_val_2
        33.8%
        22%
        76 bp
        31.1
        EPI-184_S18_L003_R1_001
        13.6%
        EPI-184_S18_L003_R1_001_val_1
        29.3%
        2.3%
        2.6%
        128.4
        25.4%
        40.8%
        26.8%
        23%
        73 bp
        20.3
        EPI-184_S18_L003_R2_001
        13.8%
        EPI-184_S18_L003_R2_001_val_2
        33.3%
        23%
        74 bp
        20.3
        EPI-185_S19_L003_R1_001
        12.2%
        EPI-185_S19_L003_R1_001_val_1
        28.9%
        2.7%
        4.7%
        57.5
        32.6%
        37.3%
        33.5%
        24%
        75 bp
        10.8
        EPI-185_S19_L003_R2_001
        12.4%
        EPI-185_S19_L003_R2_001_val_2
        36.7%
        24%
        75 bp
        10.8
        EPI-187_S20_L003_R1_001
        21.6%
        EPI-187_S20_L003_R1_001_val_1
        33.4%
        2.6%
        2.2%
        150.7
        26.5%
        42.5%
        28.2%
        23%
        75 bp
        22.3
        EPI-187_S20_L003_R2_001
        21.6%
        EPI-187_S20_L003_R2_001_val_2
        37.2%
        23%
        75 bp
        22.3
        EPI-188_S21_L003_R1_001
        26.6%
        EPI-188_S21_L003_R1_001_val_1
        28.6%
        2.6%
        2.2%
        115.9
        26.5%
        39.2%
        27.6%
        23%
        69 bp
        20.5
        EPI-188_S21_L003_R2_001
        26.6%
        EPI-188_S21_L003_R2_001_val_2
        32.8%
        23%
        69 bp
        20.5
        EPI-193_S22_L003_R1_001
        30.2%
        EPI-193_S22_L003_R1_001_val_1
        30.3%
        2.4%
        2.1%
        156.3
        23.6%
        40.0%
        24.5%
        23%
        70 bp
        25.7
        EPI-193_S22_L003_R2_001
        30.1%
        EPI-193_S22_L003_R2_001_val_2
        31.4%
        23%
        70 bp
        25.7
        EPI-194_S23_L003_R1_001
        9.9%
        EPI-194_S23_L003_R1_001_val_1
        32.0%
        2.3%
        2.1%
        203.7
        22.6%
        42.3%
        24.5%
        23%
        77 bp
        27.2
        EPI-194_S23_L003_R2_001
        10.0%
        EPI-194_S23_L003_R2_001_val_2
        36.3%
        23%
        77 bp
        27.2
        EPI-199_S24_L003_R1_001
        23.4%
        EPI-199_S24_L003_R1_001_val_1
        33.4%
        2.6%
        3.3%
        88.0
        30.3%
        39.4%
        30.3%
        24%
        72 bp
        15.4
        EPI-199_S24_L003_R2_001
        23.4%
        EPI-199_S24_L003_R2_001_val_2
        35.7%
        24%
        72 bp
        15.4
        EPI-200_S25_L003_R1_001
        24.1%
        EPI-200_S25_L003_R1_001_val_1
        32.8%
        2.6%
        2.4%
        143.4
        27.9%
        41.4%
        28.2%
        23%
        70 bp
        24.0
        EPI-200_S25_L003_R2_001
        24.2%
        EPI-200_S25_L003_R2_001_val_2
        35.6%
        23%
        70 bp
        24.0
        EPI-205_S26_L004_R1_001
        47.6%
        EPI-205_S26_L004_R1_001_val_1
        31.4%
        2.6%
        2.2%
        85.6
        25.4%
        38.2%
        27.0%
        24%
        68 bp
        15.4
        EPI-205_S26_L004_R2_001
        47.1%
        EPI-205_S26_L004_R2_001_val_2
        31.1%
        23%
        69 bp
        15.4
        EPI-206_S27_L004_R1_001
        50.2%
        EPI-206_S27_L004_R1_001_val_1
        31.3%
        2.5%
        2.2%
        128.5
        22.5%
        41.5%
        25.5%
        23%
        70 bp
        19.8
        EPI-206_S27_L004_R2_001
        49.7%
        EPI-206_S27_L004_R2_001_val_2
        31.6%
        23%
        70 bp
        19.8
        EPI-208_S28_L004_R1_001
        38.8%
        EPI-208_S28_L004_R1_001_val_1
        27.9%
        2.5%
        1.7%
        158.6
        23.1%
        38.6%
        25.3%
        23%
        68 bp
        27.2
        EPI-208_S28_L004_R2_001
        38.6%
        EPI-208_S28_L004_R2_001_val_2
        30.2%
        23%
        68 bp
        27.2
        EPI-209_S29_L004_R1_001
        17.1%
        EPI-209_S29_L004_R1_001_val_1
        31.9%
        2.4%
        1.9%
        181.3
        27.8%
        41.8%
        30.2%
        23%
        73 bp
        28.9
        EPI-209_S29_L004_R2_001
        17.2%
        EPI-209_S29_L004_R2_001_val_2
        38.4%
        23%
        73 bp
        28.9
        EPI-214_S30_L004_R1_001
        16.4%
        EPI-214_S30_L004_R1_001_val_1
        28.9%
        2.4%
        2.0%
        181.1
        24.1%
        40.0%
        25.7%
        23%
        74 bp
        28.1
        EPI-214_S30_L004_R2_001
        16.5%
        EPI-214_S30_L004_R2_001_val_2
        32.2%
        23%
        74 bp
        28.1
        EPI-215_S31_L004_R1_001
        25.4%
        EPI-215_S31_L004_R1_001_val_1
        33.7%
        2.6%
        2.2%
        126.3
        28.2%
        42.1%
        29.6%
        23%
        71 bp
        20.2
        EPI-215_S31_L004_R2_001
        25.4%
        EPI-215_S31_L004_R2_001_val_2
        36.4%
        23%
        71 bp
        20.2
        EPI-220_S32_L004_R1_001
        21.9%
        EPI-220_S32_L004_R1_001_val_1
        31.4%
        2.6%
        3.1%
        89.7
        32.3%
        41.1%
        33.2%
        23%
        73 bp
        15.3
        EPI-220_S32_L004_R2_001
        21.9%
        EPI-220_S32_L004_R2_001_val_2
        37.2%
        23%
        73 bp
        15.3
        EPI-221_S33_L004_R1_001
        41.8%
        EPI-221_S33_L004_R1_001_val_1
        31.8%
        2.7%
        2.9%
        96.7
        28.5%
        42.2%
        29.9%
        23%
        70 bp
        15.5
        EPI-221_S33_L004_R2_001
        41.5%
        EPI-221_S33_L004_R2_001_val_2
        34.6%
        23%
        71 bp
        15.5
        EPI-226_S34_L004_R1_001
        12.6%
        EPI-226_S34_L004_R1_001_val_1
        33.1%
        2.6%
        3.1%
        74.3
        26.0%
        41.7%
        28.4%
        23%
        76 bp
        11.0
        EPI-226_S34_L004_R2_001
        12.8%
        EPI-226_S34_L004_R2_001_val_2
        33.7%
        23%
        76 bp
        11.0
        EPI-227_S35_L004_R1_001
        25.0%
        EPI-227_S35_L004_R1_001_val_1
        31.6%
        2.6%
        3.1%
        86.3
        27.1%
        42.7%
        27.9%
        23%
        73 bp
        13.2
        EPI-227_S35_L004_R2_001
        25.0%
        EPI-227_S35_L004_R2_001_val_2
        32.7%
        23%
        73 bp
        13.2
        EPI-229_S36_L004_R1_001
        17.4%
        EPI-229_S36_L004_R1_001_val_1
        30.7%
        2.4%
        2.4%
        120.4
        24.7%
        40.3%
        27.6%
        24%
        72 bp
        19.3
        EPI-229_S36_L004_R2_001
        17.5%
        EPI-229_S36_L004_R2_001_val_2
        33.3%
        23%
        72 bp
        19.3
        EPI-230_S37_L004_R1_001
        21.7%
        EPI-230_S37_L004_R1_001_val_1
        30.4%
        2.2%
        1.9%
        140.0
        35.0%
        38.1%
        34.9%
        24%
        71 bp
        28.6
        EPI-230_S37_L004_R2_001
        21.7%
        EPI-230_S37_L004_R2_001_val_2
        39.1%
        23%
        71 bp
        28.6
        EPI-41_S38_L005_R1_001
        16.8%
        EPI-41_S38_L005_R1_001_val_1
        33.1%
        2.6%
        2.9%
        128.5
        36.7%
        40.4%
        35.9%
        24%
        72 bp
        24.1
        EPI-41_S38_L005_R2_001
        16.9%
        EPI-41_S38_L005_R2_001_val_2
        40.3%
        23%
        72 bp
        24.1
        EPI-42_S39_L005_R1_001
        16.1%
        EPI-42_S39_L005_R1_001_val_1
        34.7%
        2.6%
        2.1%
        167.7
        34.9%
        44.5%
        33.9%
        23%
        71 bp
        28.4
        EPI-42_S39_L005_R2_001
        16.3%
        EPI-42_S39_L005_R2_001_val_2
        42.0%
        23%
        71 bp
        28.4
        EPI-43_S40_L005_R1_001
        32.6%
        EPI-43_S40_L005_R1_001_val_1
        32.2%
        2.5%
        3.0%
        128.0
        36.5%
        44.8%
        35.0%
        23%
        72 bp
        21.9
        EPI-43_S40_L005_R2_001
        32.3%
        EPI-43_S40_L005_R2_001_val_2
        41.0%
        23%
        72 bp
        21.9
        EPI-44_S41_L005_R1_001
        66.7%
        EPI-44_S41_L005_R1_001_val_1
        34.7%
        2.6%
        2.5%
        80.0
        33.8%
        44.2%
        32.2%
        23%
        71 bp
        13.1
        EPI-44_S41_L005_R2_001
        65.5%
        EPI-44_S41_L005_R2_001_val_2
        37.7%
        23%
        71 bp
        13.1

        Bismark

        Bismark is a tool to map bisulfite converted sequence reads and determine cytosine methylation states.

        Alignment Rates

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        Deduplication

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        Strand Alignment

        All samples were run with --directional mode; alignments to complementary strands (CTOT, CTOB) were ignored.

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        Cytosine Methylation

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        M-Bias

        This plot shows the average percentage methylation and coverage across reads. See the bismark user guide for more information on how these numbers are generated.

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        Cutadapt

        Cutadapt is a tool to find and remove adapter sequences, primers, poly-Atails and other types of unwanted sequence from your high-throughput sequencing reads.

        This plot shows the number of reads with certain lengths of adapter trimmed. Obs/Exp shows the raw counts divided by the number expected due to sequencing errors. A defined peak may be related to adapter length. See the cutadapt documentation for more information on how these numbers are generated.

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


        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

        The distribution of fragment sizes (read lengths) found. See the FastQC help

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


        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.

        104 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.

        No samples found with any adapter contamination > 0.1%

        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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