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        If you use plots from MultiQC in a publication or presentation, please cite:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

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        About MultiQC

        This report was generated using MultiQC, version 1.9

        You can see a YouTube video describing how to use MultiQC reports here: https://youtu.be/qPbIlO_KWN0

        For more information about MultiQC, including other videos and extensive documentation, please visit http://multiqc.info

        You can report bugs, suggest improvements and find the source code for MultiQC on GitHub: https://github.com/ewels/MultiQC

        MultiQC is published in Bioinformatics:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

        A modular tool to aggregate results from bioinformatics analyses across many samples into a single report.

        Report generated on 2021-11-05, 09:59 based on data in: /scratch/gencore/logs/html/HF32FBGXK/merged


        General Statistics

        Showing 192/192 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HF32FBGXK_n01_5QG3108
        7.4%
        41%
        3.4
        HF32FBGXK_n01_5QG3134
        9.1%
        41%
        5.0
        HF32FBGXK_n01_5QG3150
        8.3%
        41%
        3.9
        HF32FBGXK_n01_5QG3151
        6.9%
        41%
        3.7
        HF32FBGXK_n01_5QG3153
        7.0%
        40%
        3.1
        HF32FBGXK_n01_5QG3154
        8.8%
        42%
        5.4
        HF32FBGXK_n01_5QG3157
        9.2%
        42%
        3.7
        HF32FBGXK_n01_5QG3159
        8.1%
        42%
        5.1
        HF32FBGXK_n01_5QG3160
        8.1%
        41%
        4.2
        HF32FBGXK_n01_5QG3173
        8.8%
        41%
        3.7
        HF32FBGXK_n01_5QG3181
        9.4%
        41%
        3.1
        HF32FBGXK_n01_5QG3184
        8.1%
        42%
        4.4
        HF32FBGXK_n01_5QG3185
        8.6%
        41%
        2.5
        HF32FBGXK_n01_5QG3186
        8.3%
        42%
        3.9
        HF32FBGXK_n01_5QG3186-2
        93.6%
        46%
        0.1
        HF32FBGXK_n01_5QG3193
        10.0%
        42%
        5.2
        HF32FBGXK_n01_5QG3195
        7.4%
        42%
        3.9
        HF32FBGXK_n01_5QG3195-2
        96.0%
        46%
        0.1
        HF32FBGXK_n01_5QG3199
        6.6%
        40%
        4.3
        HF32FBGXK_n01_5QG3201
        9.2%
        41%
        5.8
        HF32FBGXK_n01_5QG3210
        7.6%
        40%
        4.6
        HF32FBGXK_n01_5QG3213
        9.4%
        42%
        4.4
        HF32FBGXK_n01_5QG3221
        7.5%
        41%
        2.6
        HF32FBGXK_n01_5QG3223
        11.1%
        41%
        5.7
        HF32FBGXK_n01_5QG3224
        8.9%
        41%
        5.7
        HF32FBGXK_n01_5QG3228
        7.2%
        43%
        3.9
        HF32FBGXK_n01_5QG323
        8.4%
        34%
        3.4
        HF32FBGXK_n01_5QG3239
        7.5%
        41%
        3.1
        HF32FBGXK_n01_5QG324
        9.0%
        34%
        3.8
        HF32FBGXK_n01_5QG3241
        9.8%
        41%
        3.9
        HF32FBGXK_n01_5QG3242
        7.8%
        41%
        3.4
        HF32FBGXK_n01_5QG3244
        7.9%
        41%
        2.9
        HF32FBGXK_n01_5QG325
        8.8%
        34%
        4.5
        HF32FBGXK_n01_5QG3258
        10.6%
        41%
        1.5
        HF32FBGXK_n01_5QG326
        9.2%
        34%
        5.7
        HF32FBGXK_n01_5QG327
        12.5%
        35%
        1.5
        HF32FBGXK_n01_5QG328
        9.0%
        34%
        4.4
        HF32FBGXK_n01_5QG3282
        8.8%
        42%
        2.3
        HF32FBGXK_n01_5QG3288
        10.3%
        41%
        3.3
        HF32FBGXK_n01_5QG329
        9.8%
        35%
        6.7
        HF32FBGXK_n01_5QG3299
        8.9%
        42%
        5.6
        HF32FBGXK_n01_5QG330
        9.5%
        35%
        5.6
        HF32FBGXK_n01_5QG3307
        10.9%
        41%
        2.9
        HF32FBGXK_n01_5QG331
        10.0%
        35%
        4.5
        HF32FBGXK_n01_5QG3319
        7.8%
        42%
        5.7
        HF32FBGXK_n01_5QG332
        8.3%
        34%
        3.8
        HF32FBGXK_n01_5QG3325
        7.9%
        41%
        3.9
        HF32FBGXK_n01_5QG333
        8.7%
        34%
        5.7
        HF32FBGXK_n01_5QG3337
        9.6%
        41%
        4.7
        HF32FBGXK_n01_5QG334
        8.5%
        34%
        5.0
        HF32FBGXK_n01_5QG3341
        11.0%
        41%
        2.8
        HF32FBGXK_n01_5QG3341-2
        94.5%
        46%
        0.1
        HF32FBGXK_n01_5QG335
        8.8%
        34%
        4.2
        HF32FBGXK_n01_5QG3358
        10.5%
        41%
        4.5
        HF32FBGXK_n01_5QG3359
        9.4%
        42%
        5.6
        HF32FBGXK_n01_5QG336
        8.9%
        34%
        5.7
        HF32FBGXK_n01_5QG3362
        8.5%
        42%
        4.4
        HF32FBGXK_n01_5QG3364
        9.5%
        41%
        4.9
        HF32FBGXK_n01_5QG3367
        8.0%
        41%
        6.2
        HF32FBGXK_n01_5QG337
        8.5%
        35%
        6.3
        HF32FBGXK_n01_5QG3371
        8.4%
        41%
        4.0
        HF32FBGXK_n01_5QG3373
        9.0%
        42%
        6.1
        HF32FBGXK_n01_5QG3379
        7.8%
        41%
        4.4
        HF32FBGXK_n01_5QG338
        8.5%
        35%
        4.1
        HF32FBGXK_n01_5QG3384
        9.7%
        41%
        3.2
        HF32FBGXK_n01_5QG3386
        9.0%
        42%
        5.9
        HF32FBGXK_n01_5QG3390
        13.1%
        41%
        4.3
        HF32FBGXK_n01_5QG3391
        10.7%
        42%
        5.4
        HF32FBGXK_n01_5QG3394
        11.7%
        42%
        2.4
        HF32FBGXK_n01_5QG3394-2
        13.4%
        41%
        1.8
        HF32FBGXK_n01_5QG3398
        12.3%
        41%
        4.6
        HF32FBGXK_n01_5QG3403
        6.9%
        42%
        4.6
        HF32FBGXK_n01_5QG3403-2
        87.1%
        46%
        0.1
        HF32FBGXK_n01_5QG3410
        9.0%
        41%
        4.3
        HF32FBGXK_n01_5QG3411
        10.3%
        41%
        5.3
        HF32FBGXK_n01_5QG3412
        13.8%
        42%
        4.5
        HF32FBGXK_n01_5QG3414
        9.6%
        41%
        4.5
        HF32FBGXK_n01_5QG3429
        7.6%
        41%
        4.3
        HF32FBGXK_n01_5QG3435
        14.7%
        42%
        2.5
        HF32FBGXK_n01_5QG3443
        10.1%
        42%
        5.3
        HF32FBGXK_n01_5QG3449
        8.6%
        41%
        5.6
        HF32FBGXK_n01_5QG3453
        12.6%
        41%
        3.9
        HF32FBGXK_n01_5QG3463
        12.6%
        42%
        4.2
        HF32FBGXK_n01_5QG3464
        8.6%
        42%
        3.3
        HF32FBGXK_n01_5QG3466
        7.8%
        41%
        4.3
        HF32FBGXK_n01_5QG3475
        9.5%
        42%
        5.7
        HF32FBGXK_n01_5QG3491
        11.0%
        42%
        5.1
        HF32FBGXK_n01_5QG3492
        8.2%
        42%
        4.9
        HF32FBGXK_n01_5QG3494
        10.3%
        42%
        2.8
        HF32FBGXK_n01_5QG3499
        13.1%
        42%
        2.7
        HF32FBGXK_n01_5QG3501
        10.1%
        37%
        4.4
        HF32FBGXK_n01_5QG3503
        10.3%
        50%
        5.6
        HF32FBGXK_n01_5QG3508
        8.3%
        41%
        4.1
        HF32FBGXK_n01_5QG3510
        96.9%
        46%
        0.1
        HF32FBGXK_n01_5QG3513
        94.7%
        46%
        0.1
        HF32FBGXK_n01_undetermined
        70.9%
        43%
        15.3
        HF32FBGXK_n02_5QG3108
        7.0%
        41%
        3.4
        HF32FBGXK_n02_5QG3134
        8.5%
        42%
        5.0
        HF32FBGXK_n02_5QG3150
        7.8%
        42%
        3.9
        HF32FBGXK_n02_5QG3151
        6.6%
        42%
        3.7
        HF32FBGXK_n02_5QG3153
        6.7%
        41%
        3.1
        HF32FBGXK_n02_5QG3154
        8.3%
        43%
        5.4
        HF32FBGXK_n02_5QG3157
        8.5%
        43%
        3.7
        HF32FBGXK_n02_5QG3159
        7.7%
        42%
        5.1
        HF32FBGXK_n02_5QG3160
        7.7%
        42%
        4.2
        HF32FBGXK_n02_5QG3173
        8.2%
        42%
        3.7
        HF32FBGXK_n02_5QG3181
        8.6%
        42%
        3.1
        HF32FBGXK_n02_5QG3184
        7.4%
        43%
        4.4
        HF32FBGXK_n02_5QG3185
        7.8%
        43%
        2.5
        HF32FBGXK_n02_5QG3186
        7.6%
        43%
        3.9
        HF32FBGXK_n02_5QG3186-2
        76.0%
        79%
        0.1
        HF32FBGXK_n02_5QG3193
        9.2%
        43%
        5.2
        HF32FBGXK_n02_5QG3195
        6.8%
        43%
        3.9
        HF32FBGXK_n02_5QG3195-2
        76.3%
        79%
        0.1
        HF32FBGXK_n02_5QG3199
        6.4%
        41%
        4.3
        HF32FBGXK_n02_5QG3201
        8.7%
        42%
        5.8
        HF32FBGXK_n02_5QG3210
        7.3%
        41%
        4.6
        HF32FBGXK_n02_5QG3213
        8.7%
        43%
        4.4
        HF32FBGXK_n02_5QG3221
        6.9%
        42%
        2.6
        HF32FBGXK_n02_5QG3223
        10.8%
        42%
        5.7
        HF32FBGXK_n02_5QG3224
        8.3%
        42%
        5.7
        HF32FBGXK_n02_5QG3228
        6.7%
        43%
        3.9
        HF32FBGXK_n02_5QG323
        7.9%
        35%
        3.4
        HF32FBGXK_n02_5QG3239
        7.0%
        41%
        3.1
        HF32FBGXK_n02_5QG324
        8.2%
        36%
        3.8
        HF32FBGXK_n02_5QG3241
        9.2%
        42%
        3.9
        HF32FBGXK_n02_5QG3242
        7.3%
        41%
        3.4
        HF32FBGXK_n02_5QG3244
        7.4%
        42%
        2.9
        HF32FBGXK_n02_5QG325
        8.3%
        35%
        4.5
        HF32FBGXK_n02_5QG3258
        9.4%
        44%
        1.5
        HF32FBGXK_n02_5QG326
        8.7%
        35%
        5.7
        HF32FBGXK_n02_5QG327
        10.6%
        38%
        1.5
        HF32FBGXK_n02_5QG328
        8.2%
        35%
        4.4
        HF32FBGXK_n02_5QG3282
        8.0%
        43%
        2.3
        HF32FBGXK_n02_5QG3288
        9.2%
        43%
        3.3
        HF32FBGXK_n02_5QG329
        9.1%
        36%
        6.7
        HF32FBGXK_n02_5QG3299
        8.3%
        43%
        5.6
        HF32FBGXK_n02_5QG330
        9.0%
        36%
        5.6
        HF32FBGXK_n02_5QG3307
        9.6%
        43%
        2.9
        HF32FBGXK_n02_5QG331
        9.2%
        36%
        4.5
        HF32FBGXK_n02_5QG3319
        7.3%
        42%
        5.7
        HF32FBGXK_n02_5QG332
        7.6%
        36%
        3.8
        HF32FBGXK_n02_5QG3325
        7.6%
        41%
        3.9
        HF32FBGXK_n02_5QG333
        8.4%
        35%
        5.7
        HF32FBGXK_n02_5QG3337
        8.9%
        42%
        4.7
        HF32FBGXK_n02_5QG334
        8.1%
        35%
        5.0
        HF32FBGXK_n02_5QG3341
        9.6%
        43%
        2.8
        HF32FBGXK_n02_5QG3341-2
        73.8%
        78%
        0.1
        HF32FBGXK_n02_5QG335
        8.2%
        35%
        4.2
        HF32FBGXK_n02_5QG3358
        9.6%
        43%
        4.5
        HF32FBGXK_n02_5QG3359
        8.7%
        43%
        5.6
        HF32FBGXK_n02_5QG336
        8.6%
        35%
        5.7
        HF32FBGXK_n02_5QG3362
        7.9%
        42%
        4.4
        HF32FBGXK_n02_5QG3364
        8.9%
        42%
        4.9
        HF32FBGXK_n02_5QG3367
        7.7%
        41%
        6.2
        HF32FBGXK_n02_5QG337
        8.1%
        36%
        6.3
        HF32FBGXK_n02_5QG3371
        7.9%
        41%
        4.0
        HF32FBGXK_n02_5QG3373
        8.4%
        42%
        6.1
        HF32FBGXK_n02_5QG3379
        7.1%
        42%
        4.4
        HF32FBGXK_n02_5QG338
        8.0%
        35%
        4.1
        HF32FBGXK_n02_5QG3384
        8.6%
        43%
        3.2
        HF32FBGXK_n02_5QG3386
        8.4%
        42%
        5.9
        HF32FBGXK_n02_5QG3390
        12.5%
        42%
        4.3
        HF32FBGXK_n02_5QG3391
        9.6%
        43%
        5.4
        HF32FBGXK_n02_5QG3394
        10.5%
        44%
        2.4
        HF32FBGXK_n02_5QG3394-2
        11.5%
        44%
        1.8
        HF32FBGXK_n02_5QG3398
        11.1%
        43%
        4.6
        HF32FBGXK_n02_5QG3403
        6.6%
        43%
        4.6
        HF32FBGXK_n02_5QG3403-2
        69.9%
        76%
        0.1
        HF32FBGXK_n02_5QG3410
        8.3%
        42%
        4.3
        HF32FBGXK_n02_5QG3411
        9.7%
        42%
        5.3
        HF32FBGXK_n02_5QG3412
        12.1%
        44%
        4.5
        HF32FBGXK_n02_5QG3414
        8.9%
        42%
        4.5
        HF32FBGXK_n02_5QG3429
        7.0%
        42%
        4.3
        HF32FBGXK_n02_5QG3435
        13.1%
        46%
        2.5
        HF32FBGXK_n02_5QG3443
        9.1%
        43%
        5.3
        HF32FBGXK_n02_5QG3449
        8.1%
        42%
        5.6
        HF32FBGXK_n02_5QG3453
        11.3%
        43%
        3.9
        HF32FBGXK_n02_5QG3463
        11.3%
        44%
        4.2
        HF32FBGXK_n02_5QG3464
        7.6%
        43%
        3.3
        HF32FBGXK_n02_5QG3466
        7.2%
        42%
        4.3
        HF32FBGXK_n02_5QG3475
        8.7%
        42%
        5.7
        HF32FBGXK_n02_5QG3491
        9.9%
        44%
        5.1
        HF32FBGXK_n02_5QG3492
        7.6%
        43%
        4.9
        HF32FBGXK_n02_5QG3494
        9.3%
        43%
        2.8
        HF32FBGXK_n02_5QG3499
        11.7%
        45%
        2.7
        HF32FBGXK_n02_5QG3501
        9.3%
        39%
        4.4
        HF32FBGXK_n02_5QG3503
        9.4%
        51%
        5.6
        HF32FBGXK_n02_5QG3508
        7.7%
        42%
        4.1
        HF32FBGXK_n02_5QG3510
        81.3%
        83%
        0.1
        HF32FBGXK_n02_5QG3513
        76.4%
        80%
        0.1
        HF32FBGXK_n02_undetermined
        70.2%
        43%
        15.3

        Demultiplexing Report


        Total Read Count: Total number of PF (Passing Filter) reads in this library.
        Portion: The proportion of reads that represent the individual library in the entire Library Pool.

        Showing 96/96 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        15289323
        3.8
        5QG323
        3409243
        0.9
        5QG331
        4535711
        1.1
        5QG3108
        3429758
        0.9
        5QG3160
        4195979
        1.1
        5QG3199
        4295368
        1.1
        5QG3239
        3080516
        0.8
        5QG3307
        2926006
        0.7
        5QG3364
        4858593
        1.2
        5QG3391
        5439169
        1.4
        5QG3429
        4321521
        1.1
        5QG3475
        5711778
        1.4
        5QG3510
        92783
        0.0
        5QG324
        3770170
        0.9
        5QG332
        3756109
        0.9
        5QG3134
        5047510
        1.3
        5QG3173
        3695279
        0.9
        5QG3201
        5777559
        1.5
        5QG3241
        3850292
        1.0
        5QG3319
        5710948
        1.4
        5QG3367
        6208812
        1.6
        5QG3394
        2362750
        0.6
        5QG3435
        2544956
        0.6
        5QG3491
        5115820
        1.3
        5QG3513
        109269
        0.0
        5QG325
        4535782
        1.1
        5QG333
        5726617
        1.4
        5QG3150
        3937454
        1.0
        5QG3181
        3055084
        0.8
        5QG3210
        4551168
        1.1
        5QG3242
        3374400
        0.8
        5QG3325
        3926403
        1.0
        5QG3371
        3996699
        1.0
        5QG3398
        4596594
        1.2
        5QG3443
        5267405
        1.3
        5QG3492
        4881490
        1.2
        5QG3186-2
        60651
        0.0
        5QG326
        5691106
        1.4
        5QG334
        5028088
        1.3
        5QG3151
        3680145
        0.9
        5QG3184
        4382868
        1.1
        5QG3213
        4390802
        1.1
        5QG3244
        2866585
        0.7
        5QG3337
        4707522
        1.2
        5QG3373
        6107342
        1.5
        5QG3403
        4587881
        1.2
        5QG3449
        5580845
        1.4
        5QG3494
        2846899
        0.7
        5QG3195-2
        65908
        0.0
        5QG327
        1500640
        0.4
        5QG335
        4151544
        1.0
        5QG3153
        3079642
        0.8
        5QG3185
        2481503
        0.6
        5QG3221
        2643016
        0.7
        5QG3258
        1512686
        0.4
        5QG3341
        2773444
        0.7
        5QG3379
        4412060
        1.1
        5QG3410
        4319279
        1.1
        5QG3453
        3878296
        1.0
        5QG3499
        2743960
        0.7
        5QG3341-2
        83941
        0.0
        5QG328
        4374160
        1.1
        5QG336
        5725066
        1.4
        5QG3154
        5374346
        1.4
        5QG3186
        3893947
        1.0
        5QG3223
        5673778
        1.4
        5QG3282
        2253528
        0.6
        5QG3358
        4541242
        1.1
        5QG3384
        3188697
        0.8
        5QG3411
        5301422
        1.3
        5QG3463
        4221001
        1.1
        5QG3501
        4358385
        1.1
        5QG3394-2
        1755160
        0.4
        5QG329
        6706149
        1.7
        5QG337
        6296541
        1.6
        5QG3157
        3727499
        0.9
        5QG3193
        5184634
        1.3
        5QG3224
        5739763
        1.4
        5QG3288
        3280907
        0.8
        5QG3359
        5565026
        1.4
        5QG3386
        5851310
        1.5
        5QG3412
        4485936
        1.1
        5QG3464
        3294800
        0.8
        5QG3503
        5607818
        1.4
        5QG3403-2
        96619
        0.0
        5QG330
        5585588
        1.4
        5QG338
        4059934
        1.0
        5QG3159
        5095945
        1.3
        5QG3195
        3857343
        1.0
        5QG3228
        3866188
        1.0
        5QG3299
        5648137
        1.4
        5QG3362
        4382515
        1.1
        5QG3390
        4296585
        1.1
        5QG3414
        4542430
        1.1
        5QG3466
        4292243
        1.1
        5QG3508
        4145692
        1.0

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        4.0
        418666488
        397231305
        3.9
        2.7

        Barcodes of Undetermined Reads


        We have determined the barcodes of your undetermined reads (reads containing a barcode that you did not encode in your metadata). Here are the top 20 barcodes belonging to the undetermined reads. The full list is available here.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGGGG
        11938393.0
        78.1
        NNNNNNNN
        132611.0
        0.9
        GGGGGGGC
        25268.0
        0.2
        GGGGGGGT
        14990.0
        0.1
        GAACGTTA
        13350.0
        0.1
        GGGGGGTG
        10923.0
        0.1
        GGGGGGCC
        10540.0
        0.1
        GGGGGGCG
        9888.0
        0.1
        AACGGTTA
        9372.0
        0.1
        GGGGGCCC
        9146.0
        0.1
        GTGGGGGG
        8702.0
        0.1
        GGGGGCGG
        8211.0
        0.1
        GNNNNNNN
        8016.0
        0.1
        GCCTAACA
        7945.0
        0.1
        CNNNNNNN
        7610.0
        0.1
        TGCGTACA
        7546.0
        0.1
        GCGGGGGG
        7541.0
        0.1
        GAAGGTTA
        7489.0
        0.1
        GCCTTACA
        7409.0
        0.1
        GACGGTTA
        7005.0
        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

        All samples have sequences of a single length (76bp).

        Sequence Duplication Levels

        The relative level of duplication found for every sequence.

        From the FastQC Help:

        In a diverse library most sequences will occur only once in the final set. A low level of duplication may indicate a very high level of coverage of the target sequence, but a high level of duplication is more likely to indicate some kind of enrichment bias (eg PCR over amplification). This graph shows the degree of duplication for every sequence in a library: the relative number of sequences with different degrees of duplication.

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        In a properly diverse library most sequences should fall into the far left of the plot in both the red and blue lines. A general level of enrichment, indicating broad oversequencing in the library will tend to flatten the lines, lowering the low end and generally raising other categories. More specific enrichments of subsets, or the presence of low complexity contaminants will tend to produce spikes towards the right of the plot.

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


        Overrepresented sequences

        The total amount of overrepresented sequences found in each library.

        FastQC calculates and lists overrepresented sequences in FastQ files. It would not be possible to show this for all samples in a MultiQC report, so instead this plot shows the number of sequences categorized as over represented.

        Sometimes, a single sequence may account for a large number of reads in a dataset. To show this, the bars are split into two: the first shows the overrepresented reads that come from the single most common sequence. The second shows the total count from all remaining overrepresented sequences.

        From the FastQC Help:

        A normal high-throughput library will contain a diverse set of sequences, with no individual sequence making up a tiny fraction of the whole. Finding that a single sequence is very overrepresented in the set either means that it is highly biologically significant, or indicates that the library is contaminated, or not as diverse as you expected.

        FastQC lists all of the sequences which make up more than 0.1% of the total. To conserve memory only sequences which appear in the first 100,000 sequences are tracked to the end of the file. It is therefore possible that a sequence which is overrepresented but doesn't appear at the start of the file for some reason could be missed by this module.

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


        Adapter Content

        The cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position.

        Note that only samples with ≥ 0.1% adapter contamination are shown.

        There may be several lines per sample, as one is shown for each adapter detected in the file.

        From the FastQC Help:

        The plot shows a cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position. Once a sequence has been seen in a read it is counted as being present right through to the end of the read so the percentages you see will only increase as the read length goes on.

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


        Status Checks

        Status for each FastQC section showing whether results seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        FastQC assigns a status for each section of the report. These give a quick evaluation of whether the results of the analysis seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        It is important to stress that although the analysis results appear to give a pass/fail result, these evaluations must be taken in the context of what you expect from your library. A 'normal' sample as far as FastQC is concerned is random and diverse. Some experiments may be expected to produce libraries which are biased in particular ways. You should treat the summary evaluations therefore as pointers to where you should concentrate your attention and understand why your library may not look random and diverse.

        Specific guidance on how to interpret the output of each module can be found in the relevant report section, or in the FastQC help.

        In this heatmap, we summarise all of these into a single heatmap for a quick overview. Note that not all FastQC sections have plots in MultiQC reports, but all status checks are shown in this heatmap.

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