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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 1.11

        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 2023-10-31, 17:06 based on data in: /work_home/orue/PROJECTS/LEBANESEWHEATSOURDOUGH/16S/RACHELLE/FASTQC


        General Statistics

        Showing 276/276 rows and 4/5 columns.
        Sample Name% Dups% GCLengthM Seqs
        B1A_R1
        55.1%
        51%
        300 bp
        0.1
        B1A_R2
        83.2%
        50%
        300 bp
        0.1
        B1B_R1
        54.5%
        49%
        290 bp
        0.1
        B1B_R2
        77.9%
        48%
        291 bp
        0.1
        B2A_R1
        49.0%
        53%
        301 bp
        0.1
        B2A_R2
        74.5%
        52%
        300 bp
        0.1
        B2B_R1
        37.1%
        52%
        292 bp
        0.1
        B2B_R2
        67.5%
        51%
        291 bp
        0.1
        B3A_R1
        46.1%
        53%
        300 bp
        0.0
        B3A_R2
        74.8%
        52%
        299 bp
        0.0
        B3F1_R1
        55.1%
        53%
        299 bp
        0.1
        B3F1_R2
        82.1%
        52%
        298 bp
        0.1
        B4-2A_R1
        60.4%
        53%
        300 bp
        0.1
        B4-2A_R2
        81.9%
        52%
        300 bp
        0.1
        B4-2B_R1
        46.4%
        53%
        288 bp
        0.0
        B4-2B_R2
        66.6%
        52%
        287 bp
        0.0
        B4-2C_R1
        29.3%
        53%
        261 bp
        0.0
        B4-2C_R2
        48.0%
        53%
        261 bp
        0.0
        B4-2F1-1_R1
        48.9%
        53%
        288 bp
        0.0
        B4-2F1-1_R2
        71.0%
        52%
        288 bp
        0.0
        B4-2F1-2_R1
        60.4%
        53%
        296 bp
        0.1
        B4-2F1-2_R2
        80.5%
        52%
        296 bp
        0.1
        B4-2F2_R1
        57.4%
        53%
        299 bp
        0.0
        B4-2F2_R2
        78.9%
        52%
        298 bp
        0.0
        B4A-1_R1
        55.7%
        53%
        279 bp
        0.0
        B4A-1_R2
        79.0%
        52%
        279 bp
        0.0
        B4A-2_R1
        68.9%
        53%
        301 bp
        0.1
        B4A-2_R2
        89.9%
        52%
        300 bp
        0.1
        B4B_R1
        42.1%
        52%
        290 bp
        0.0
        B4B_R2
        71.2%
        51%
        290 bp
        0.0
        B4C_R1
        25.6%
        53%
        251 bp
        0.0
        B4C_R2
        39.0%
        53%
        251 bp
        0.0
        B4F1_R1
        57.2%
        53%
        294 bp
        0.1
        B4F1_R2
        80.0%
        52%
        294 bp
        0.1
        B5A_R1
        60.1%
        53%
        300 bp
        0.1
        B5A_R2
        83.4%
        52%
        299 bp
        0.1
        B5B_R1
        39.8%
        53%
        248 bp
        0.1
        B5B_R2
        57.4%
        52%
        247 bp
        0.1
        B5F1_R1
        55.2%
        53%
        291 bp
        0.1
        B5F1_R2
        77.7%
        52%
        290 bp
        0.1
        B5F2_R1
        56.9%
        53%
        290 bp
        0.1
        B5F2_R2
        77.8%
        52%
        290 bp
        0.1
        Blanc-PCR1_R1
        2.0%
        50%
        53 bp
        0.0
        Blanc-PCR1_R2
        1.7%
        49%
        55 bp
        0.0
        Blanc-PCR2_R1
        2.3%
        50%
        52 bp
        0.0
        Blanc-PCR2_R2
        1.7%
        50%
        54 bp
        0.0
        Blanc-PCR3_R1
        2.9%
        50%
        93 bp
        0.0
        Blanc-PCR3_R2
        2.9%
        50%
        93 bp
        0.0
        Blanc-PCR4_R1
        0.9%
        50%
        57 bp
        0.0
        Blanc-PCR4_R2
        0.8%
        50%
        58 bp
        0.0
        Blanc-d-ext-X10_R1
        2.0%
        50%
        55 bp
        0.0
        Blanc-d-ext-X10_R2
        1.3%
        50%
        57 bp
        0.0
        Blanc-d-ext-X11_R1
        2.3%
        50%
        52 bp
        0.0
        Blanc-d-ext-X11_R2
        1.6%
        50%
        54 bp
        0.0
        Blanc-d-ext-X2_R1
        5.2%
        50%
        69 bp
        0.0
        Blanc-d-ext-X2_R2
        4.2%
        50%
        70 bp
        0.0
        Blanc-d-ext-X3_R1
        1.8%
        50%
        52 bp
        0.0
        Blanc-d-ext-X3_R2
        1.2%
        50%
        54 bp
        0.0
        Blanc-d-ext-X4_R1
        3.2%
        51%
        76 bp
        0.0
        Blanc-d-ext-X4_R2
        3.0%
        51%
        79 bp
        0.0
        Blanc-d-ext-X5-1_R1
        0.4%
        55%
        298 bp
        0.0
        Blanc-d-ext-X5-1_R2
        2.4%
        55%
        297 bp
        0.0
        Blanc-d-ext-X5-2_R1
        17.8%
        54%
        221 bp
        0.0
        Blanc-d-ext-X5-2_R2
        31.1%
        53%
        222 bp
        0.0
        Blanc-d-ext-X6_R1
        10.2%
        52%
        135 bp
        0.0
        Blanc-d-ext-X6_R2
        15.9%
        51%
        137 bp
        0.0
        Blanc-d-ext-X7_R1
        4.2%
        51%
        89 bp
        0.0
        Blanc-d-ext-X7_R2
        5.6%
        51%
        91 bp
        0.0
        Blanc-d-ext-X8_R1
        11.1%
        52%
        155 bp
        0.0
        Blanc-d-ext-X8_R2
        18.3%
        51%
        157 bp
        0.0
        Blanc-d-ext-X9_R1
        2.5%
        50%
        61 bp
        0.0
        Blanc-d-ext-X9_R2
        1.7%
        50%
        65 bp
        0.0
        FB4A_R1
        59.4%
        53%
        300 bp
        0.0
        FB4A_R2
        85.1%
        52%
        299 bp
        0.0
        FB4F1_R1
        64.2%
        53%
        296 bp
        0.1
        FB4F1_R2
        83.7%
        52%
        296 bp
        0.1
        FB4F2_R1
        54.1%
        52%
        294 bp
        0.1
        FB4F2_R2
        82.4%
        51%
        294 bp
        0.1
        FB5A_R1
        58.4%
        53%
        300 bp
        0.1
        FB5A_R2
        81.8%
        52%
        299 bp
        0.1
        FJ4A_R1
        45.7%
        53%
        300 bp
        0.1
        FJ4A_R2
        72.5%
        51%
        300 bp
        0.1
        FS3A_R1
        46.1%
        51%
        300 bp
        0.0
        FS3A_R2
        78.2%
        51%
        299 bp
        0.0
        J1A_R1
        61.5%
        53%
        301 bp
        0.1
        J1A_R2
        84.7%
        52%
        300 bp
        0.1
        J1B_R1
        41.2%
        54%
        261 bp
        0.1
        J1B_R2
        57.6%
        53%
        261 bp
        0.1
        J1C_R1
        32.1%
        54%
        246 bp
        0.0
        J1C_R2
        45.4%
        54%
        246 bp
        0.0
        J2-2A_R1
        67.2%
        53%
        301 bp
        0.1
        J2-2A_R2
        87.4%
        52%
        300 bp
        0.1
        J2-2B_R1
        39.1%
        52%
        283 bp
        0.0
        J2-2B_R2
        67.3%
        52%
        283 bp
        0.0
        J2-2F1_R1
        60.1%
        53%
        299 bp
        0.1
        J2-2F1_R2
        80.5%
        52%
        298 bp
        0.1
        J2-2F2-1_R1
        50.4%
        52%
        294 bp
        0.0
        J2-2F2-1_R2
        73.5%
        52%
        294 bp
        0.0
        J2-2F2-2_R1
        61.8%
        52%
        300 bp
        0.1
        J2-2F2-2_R2
        82.4%
        52%
        299 bp
        0.1
        J2A_R1
        62.0%
        53%
        301 bp
        0.1
        J2A_R2
        84.8%
        52%
        300 bp
        0.1
        J2B_R1
        50.4%
        50%
        298 bp
        0.1
        J2B_R2
        80.1%
        49%
        299 bp
        0.1
        J2C_R1
        38.0%
        53%
        261 bp
        0.1
        J2C_R2
        56.2%
        53%
        261 bp
        0.1
        J2F1_R1
        56.3%
        52%
        299 bp
        0.1
        J2F1_R2
        80.8%
        51%
        298 bp
        0.1
        J3A_R1
        46.6%
        53%
        301 bp
        0.0
        J3A_R2
        76.7%
        52%
        300 bp
        0.0
        J3B_R1
        48.2%
        50%
        297 bp
        0.1
        J3B_R2
        78.7%
        49%
        297 bp
        0.1
        J3C_R1
        21.9%
        54%
        215 bp
        0.0
        J3C_R2
        33.4%
        53%
        215 bp
        0.0
        J4A_R1
        50.0%
        55%
        300 bp
        0.1
        J4A_R2
        78.7%
        53%
        300 bp
        0.1
        J4B_R1
        37.7%
        55%
        274 bp
        0.1
        J4B_R2
        58.9%
        53%
        273 bp
        0.1
        J4C_R1
        37.5%
        54%
        248 bp
        0.1
        J4C_R2
        51.3%
        54%
        248 bp
        0.1
        J4F1_R1
        36.4%
        54%
        283 bp
        0.1
        J4F1_R2
        58.0%
        53%
        283 bp
        0.1
        J5-2A_R1
        50.8%
        53%
        295 bp
        0.0
        J5-2A_R2
        77.1%
        52%
        293 bp
        0.0
        J5-2B_R1
        31.4%
        52%
        282 bp
        0.0
        J5-2B_R2
        58.2%
        51%
        282 bp
        0.0
        J5-2F1_R1
        57.3%
        53%
        299 bp
        0.1
        J5-2F1_R2
        83.2%
        52%
        298 bp
        0.1
        J5-2F2_R1
        64.8%
        53%
        300 bp
        0.1
        J5-2F2_R2
        87.9%
        52%
        299 bp
        0.1
        J5A_R1
        49.7%
        53%
        296 bp
        0.0
        J5A_R2
        76.6%
        52%
        295 bp
        0.0
        J5B_R1
        57.0%
        53%
        295 bp
        0.0
        J5B_R2
        80.7%
        52%
        294 bp
        0.0
        J5C_R1
        30.0%
        54%
        242 bp
        0.0
        J5C_R2
        44.4%
        53%
        242 bp
        0.0
        K1A_R1
        58.1%
        53%
        300 bp
        0.1
        K1A_R2
        80.3%
        52%
        300 bp
        0.1
        K1B_R1
        49.4%
        52%
        296 bp
        0.0
        K1B_R2
        82.6%
        51%
        296 bp
        0.0
        K2A_R1
        58.4%
        53%
        300 bp
        0.1
        K2A_R2
        79.4%
        52%
        299 bp
        0.1
        K2B_R1
        50.2%
        51%
        297 bp
        0.1
        K2B_R2
        76.2%
        51%
        296 bp
        0.1
        K3A_R1
        46.3%
        52%
        299 bp
        0.0
        K3A_R2
        81.3%
        51%
        299 bp
        0.0
        K3B_R1
        49.6%
        52%
        299 bp
        0.0
        K3B_R2
        84.4%
        51%
        299 bp
        0.0
        K3C_R1
        30.7%
        54%
        266 bp
        0.1
        K3C_R2
        48.4%
        54%
        266 bp
        0.1
        K4B_R1
        47.0%
        53%
        288 bp
        0.1
        K4B_R2
        73.3%
        52%
        288 bp
        0.1
        K5A_R1
        60.6%
        53%
        300 bp
        0.1
        K5A_R2
        86.6%
        52%
        299 bp
        0.1
        K5B_R1
        40.8%
        54%
        274 bp
        0.1
        K5B_R2
        63.8%
        53%
        273 bp
        0.1
        K5F1_R1
        61.8%
        53%
        299 bp
        0.1
        K5F1_R2
        85.4%
        52%
        298 bp
        0.1
        M1-2B1_R1
        43.2%
        51%
        298 bp
        0.1
        M1-2B1_R2
        76.6%
        51%
        297 bp
        0.1
        M1-2B2_R1
        41.7%
        51%
        299 bp
        0.0
        M1-2B2_R2
        75.3%
        51%
        299 bp
        0.0
        M1-2B3_R1
        42.3%
        51%
        299 bp
        0.0
        M1-2B3_R2
        75.3%
        51%
        299 bp
        0.0
        M1-2C_R1
        33.6%
        55%
        254 bp
        0.1
        M1-2C_R2
        48.3%
        54%
        254 bp
        0.1
        M1A_R1
        46.1%
        52%
        299 bp
        0.0
        M1A_R2
        84.0%
        51%
        299 bp
        0.0
        M1B_R1
        55.0%
        52%
        300 bp
        0.1
        M1B_R2
        86.1%
        51%
        300 bp
        0.1
        M1C_R1
        22.1%
        54%
        214 bp
        0.0
        M1C_R2
        32.7%
        53%
        214 bp
        0.0
        M1F1_R1
        56.8%
        53%
        299 bp
        0.1
        M1F1_R2
        83.7%
        52%
        299 bp
        0.1
        MOCK-bacterie4_R1
        70.1%
        51%
        301 bp
        0.1
        MOCK-bacterie4_R2
        86.9%
        49%
        300 bp
        0.1
        MOCK-bacterie5_R1
        66.3%
        51%
        301 bp
        0.1
        MOCK-bacterie5_R2
        84.4%
        49%
        300 bp
        0.1
        Mock-bacterie1_R1
        63.0%
        50%
        300 bp
        0.1
        Mock-bacterie1_R2
        82.4%
        49%
        300 bp
        0.1
        Mock-bacterie2_R1
        63.5%
        50%
        300 bp
        0.1
        Mock-bacterie2_R2
        82.8%
        49%
        300 bp
        0.1
        Mock-bacterie3_R1
        53.8%
        51%
        298 bp
        0.0
        Mock-bacterie3_R2
        73.5%
        49%
        297 bp
        0.0
        N1A_R1
        57.6%
        53%
        300 bp
        0.1
        N1A_R2
        83.9%
        52%
        300 bp
        0.1
        N1F1_R1
        60.9%
        52%
        300 bp
        0.1
        N1F1_R2
        85.3%
        51%
        299 bp
        0.1
        N2A_R1
        65.2%
        53%
        301 bp
        0.1
        N2A_R2
        87.6%
        52%
        300 bp
        0.1
        N2B_R1
        35.6%
        52%
        277 bp
        0.0
        N2B_R2
        59.8%
        52%
        276 bp
        0.0
        N3A_R1
        60.8%
        53%
        301 bp
        0.1
        N3A_R2
        84.5%
        52%
        300 bp
        0.1
        N3B_R1
        48.6%
        53%
        293 bp
        0.0
        N3B_R2
        75.4%
        52%
        293 bp
        0.0
        N3C_R1
        28.1%
        53%
        254 bp
        0.0
        N3C_R2
        44.2%
        53%
        253 bp
        0.0
        N4-2A_R1
        61.6%
        53%
        300 bp
        0.1
        N4-2A_R2
        85.0%
        52%
        300 bp
        0.1
        N4-2B_R1
        46.7%
        49%
        293 bp
        0.0
        N4-2B_R2
        75.2%
        49%
        294 bp
        0.0
        N4-2F2_R1
        51.2%
        51%
        298 bp
        0.1
        N4-2F2_R2
        81.1%
        50%
        298 bp
        0.1
        N4A_R1
        51.6%
        53%
        300 bp
        0.1
        N4A_R2
        78.4%
        52%
        299 bp
        0.1
        N4B_R1
        43.4%
        51%
        292 bp
        0.0
        N4B_R2
        74.7%
        51%
        292 bp
        0.0
        N5A_R1
        56.6%
        53%
        301 bp
        0.1
        N5A_R2
        79.7%
        52%
        300 bp
        0.1
        N5C_R1
        38.8%
        54%
        256 bp
        0.1
        N5C_R2
        53.1%
        54%
        255 bp
        0.1
        N5F2_R1
        53.5%
        53%
        298 bp
        0.1
        N5F2_R2
        76.8%
        52%
        298 bp
        0.1
        S1-2A_R1
        60.6%
        53%
        300 bp
        0.0
        S1-2A_R2
        79.8%
        52%
        300 bp
        0.0
        S1-2B_R1
        59.4%
        53%
        296 bp
        0.1
        S1-2B_R2
        84.3%
        52%
        296 bp
        0.1
        S1-2C_R1
        27.4%
        53%
        242 bp
        0.1
        S1-2C_R2
        43.1%
        53%
        241 bp
        0.1
        S1-2F1_R1
        55.5%
        53%
        296 bp
        0.0
        S1-2F1_R2
        80.5%
        52%
        296 bp
        0.0
        S1-2F2_R1
        66.4%
        53%
        297 bp
        0.1
        S1-2F2_R2
        85.3%
        52%
        297 bp
        0.1
        S1A_R1
        61.6%
        53%
        301 bp
        0.1
        S1A_R2
        84.9%
        52%
        300 bp
        0.1
        S1B-1_R1
        36.7%
        52%
        271 bp
        0.0
        S1B-1_R2
        64.8%
        51%
        271 bp
        0.0
        S1B-2_R1
        47.3%
        52%
        298 bp
        0.1
        S1B-2_R2
        79.7%
        51%
        298 bp
        0.1
        S2A_R1
        56.0%
        53%
        296 bp
        0.1
        S2A_R2
        81.5%
        52%
        296 bp
        0.1
        S2B_R1
        49.1%
        50%
        290 bp
        0.1
        S2B_R2
        72.9%
        50%
        290 bp
        0.1
        S2C_R1
        31.6%
        54%
        257 bp
        0.1
        S2C_R2
        47.2%
        54%
        257 bp
        0.1
        S2F1_R1
        61.9%
        53%
        297 bp
        0.1
        S2F1_R2
        79.8%
        52%
        296 bp
        0.1
        S3A_R1
        59.4%
        53%
        301 bp
        0.0
        S3A_R2
        84.8%
        52%
        300 bp
        0.0
        S3B_R1
        35.4%
        52%
        291 bp
        0.0
        S3B_R2
        72.2%
        51%
        291 bp
        0.0
        S3C_R1
        31.0%
        53%
        259 bp
        0.1
        S3C_R2
        48.9%
        52%
        259 bp
        0.1
        S3F2_R1
        58.5%
        53%
        300 bp
        0.1
        S3F2_R2
        87.4%
        52%
        299 bp
        0.1
        S4A_R1
        55.2%
        53%
        299 bp
        0.0
        S4A_R2
        78.1%
        52%
        299 bp
        0.0
        S4B_R1
        35.5%
        52%
        274 bp
        0.0
        S4B_R2
        64.4%
        52%
        274 bp
        0.0
        S4F1-1_R1
        48.3%
        53%
        278 bp
        0.0
        S4F1-1_R2
        68.5%
        52%
        277 bp
        0.0
        S4F1-2_R1
        56.8%
        53%
        293 bp
        0.1
        S4F1-2_R2
        76.1%
        52%
        292 bp
        0.1
        S4F2_R1
        56.0%
        53%
        297 bp
        0.1
        S4F2_R2
        81.2%
        52%
        297 bp
        0.1
        S5A_R1
        63.4%
        52%
        301 bp
        0.1
        S5A_R2
        85.0%
        51%
        300 bp
        0.1
        S5B_R1
        32.3%
        52%
        289 bp
        0.0
        S5B_R2
        59.3%
        51%
        289 bp
        0.0
        S5F1_R1
        62.8%
        53%
        298 bp
        0.1
        S5F1_R2
        83.3%
        52%
        297 bp
        0.1
        T1A-a_R1
        60.6%
        53%
        300 bp
        0.1
        T1A-a_R2
        84.9%
        52%
        299 bp
        0.1
        T1A-b_R1
        65.8%
        53%
        301 bp
        0.1
        T1A-b_R2
        84.5%
        52%
        300 bp
        0.1
        T1F1_R1
        59.7%
        53%
        299 bp
        0.1
        T1F1_R2
        79.4%
        52%
        298 bp
        0.1
        T2A_R1
        53.2%
        53%
        301 bp
        0.1
        T2A_R2
        78.6%
        52%
        300 bp
        0.1
        T2B_R1
        49.3%
        53%
        282 bp
        0.0
        T2B_R2
        70.9%
        52%
        282 bp
        0.0
        T2F1_R1
        60.0%
        53%
        299 bp
        0.1
        T2F1_R2
        80.7%
        52%
        299 bp
        0.1
        T2F2_R1
        60.8%
        54%
        299 bp
        0.1
        T2F2_R2
        81.2%
        53%
        298 bp
        0.1

        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.

        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.

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        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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