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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 2021-07-20, 11:34 based on data in: /scratch/gencore/logs/html/HGV55BGXJ/merged


        General Statistics

        Showing 190/190 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HGV55BGXJ_n01_4QG3117
        10.3%
        42%
        5.0
        HGV55BGXJ_n01_4QG3120
        10.1%
        42%
        3.7
        HGV55BGXJ_n01_4QG3126
        13.8%
        42%
        4.8
        HGV55BGXJ_n01_4QG3129
        10.0%
        42%
        6.3
        HGV55BGXJ_n01_4QG3155
        11.4%
        42%
        3.9
        HGV55BGXJ_n01_4QG3161
        9.5%
        41%
        3.7
        HGV55BGXJ_n01_4QG3172
        10.3%
        42%
        5.6
        HGV55BGXJ_n01_4QG3189
        11.5%
        42%
        6.9
        HGV55BGXJ_n01_4QG3197
        8.9%
        42%
        6.6
        HGV55BGXJ_n01_4QG3205
        11.6%
        42%
        6.3
        HGV55BGXJ_n01_4QG3209
        13.7%
        42%
        4.2
        HGV55BGXJ_n01_4QG3225
        10.2%
        42%
        6.0
        HGV55BGXJ_n01_4QG3229
        9.5%
        42%
        6.4
        HGV55BGXJ_n01_4QG3232
        8.9%
        42%
        5.8
        HGV55BGXJ_n01_4QG3248
        7.7%
        41%
        3.4
        HGV55BGXJ_n01_4QG3254
        11.0%
        42%
        5.9
        HGV55BGXJ_n01_4QG3257
        11.3%
        42%
        6.0
        HGV55BGXJ_n01_4QG3259
        10.5%
        41%
        2.6
        HGV55BGXJ_n01_4QG3262
        12.7%
        42%
        4.2
        HGV55BGXJ_n01_4QG3263
        12.5%
        42%
        4.4
        HGV55BGXJ_n01_4QG3264
        11.6%
        41%
        1.7
        HGV55BGXJ_n01_4QG3267
        9.6%
        42%
        2.1
        HGV55BGXJ_n01_4QG3270
        11.7%
        42%
        2.6
        HGV55BGXJ_n01_4QG3272
        9.3%
        41%
        2.5
        HGV55BGXJ_n01_4QG3273
        11.8%
        42%
        4.4
        HGV55BGXJ_n01_4QG3274
        18.5%
        42%
        2.0
        HGV55BGXJ_n01_4QG3289
        15.8%
        42%
        6.1
        HGV55BGXJ_n01_4QG3292
        11.0%
        42%
        5.6
        HGV55BGXJ_n01_4QG3297
        11.3%
        42%
        1.8
        HGV55BGXJ_n01_4QG3300
        9.7%
        42%
        5.1
        HGV55BGXJ_n01_4QG3301
        12.4%
        42%
        5.1
        HGV55BGXJ_n01_4QG3303
        9.9%
        42%
        3.6
        HGV55BGXJ_n01_4QG3307
        17.0%
        42%
        1.6
        HGV55BGXJ_n01_4QG3316
        11.4%
        42%
        2.6
        HGV55BGXJ_n01_4QG3320
        12.6%
        43%
        5.2
        HGV55BGXJ_n01_4QG3327
        22.3%
        43%
        3.0
        HGV55BGXJ_n01_4QG3332
        15.4%
        42%
        1.9
        HGV55BGXJ_n01_4QG3339
        16.1%
        42%
        4.4
        HGV55BGXJ_n01_4QG3345
        10.9%
        42%
        3.7
        HGV55BGXJ_n01_4QG3346
        13.2%
        42%
        5.1
        HGV55BGXJ_n01_4QG3349
        12.1%
        42%
        4.4
        HGV55BGXJ_n01_4QG3350
        13.7%
        42%
        7.7
        HGV55BGXJ_n01_4QG3352
        14.1%
        42%
        4.9
        HGV55BGXJ_n01_4QG3354
        11.8%
        42%
        4.9
        HGV55BGXJ_n01_4QG3355
        11.8%
        42%
        4.5
        HGV55BGXJ_n01_4QG3356
        16.3%
        42%
        2.7
        HGV55BGXJ_n01_4QG3357
        12.8%
        42%
        3.4
        HGV55BGXJ_n01_4QG3358
        13.9%
        42%
        1.3
        HGV55BGXJ_n01_4QG3360
        15.2%
        42%
        7.4
        HGV55BGXJ_n01_4QG3363
        12.4%
        42%
        2.5
        HGV55BGXJ_n01_4QG3365
        14.3%
        42%
        4.7
        HGV55BGXJ_n01_4QG3366
        15.1%
        42%
        4.4
        HGV55BGXJ_n01_4QG3367
        14.2%
        42%
        1.7
        HGV55BGXJ_n01_4QG3370
        14.1%
        42%
        2.2
        HGV55BGXJ_n01_4QG3372
        14.6%
        42%
        5.3
        HGV55BGXJ_n01_4QG3375
        12.2%
        42%
        1.7
        HGV55BGXJ_n01_4QG3387
        13.3%
        42%
        5.6
        HGV55BGXJ_n01_4QG3395
        16.7%
        42%
        2.4
        HGV55BGXJ_n01_4QG3400
        12.6%
        42%
        5.6
        HGV55BGXJ_n01_4QG3401
        13.8%
        42%
        3.7
        HGV55BGXJ_n01_4QG3404
        13.0%
        42%
        4.6
        HGV55BGXJ_n01_4QG3405
        13.9%
        42%
        3.7
        HGV55BGXJ_n01_4QG3407
        13.7%
        42%
        4.5
        HGV55BGXJ_n01_4QG3409
        11.4%
        42%
        3.8
        HGV55BGXJ_n01_4QG3412
        22.5%
        44%
        0.0
        HGV55BGXJ_n01_4QG3415
        9.9%
        42%
        3.6
        HGV55BGXJ_n01_4QG3421
        9.2%
        42%
        2.6
        HGV55BGXJ_n01_4QG3422
        12.0%
        42%
        4.9
        HGV55BGXJ_n01_4QG3425
        12.8%
        42%
        3.4
        HGV55BGXJ_n01_4QG3426
        11.1%
        42%
        3.1
        HGV55BGXJ_n01_4QG3430
        10.7%
        42%
        5.5
        HGV55BGXJ_n01_4QG3432
        10.4%
        42%
        4.2
        HGV55BGXJ_n01_4QG3436
        11.9%
        42%
        4.7
        HGV55BGXJ_n01_4QG3437
        12.6%
        42%
        3.3
        HGV55BGXJ_n01_4QG3441
        10.3%
        42%
        3.2
        HGV55BGXJ_n01_4QG3442
        15.8%
        42%
        2.6
        HGV55BGXJ_n01_4QG3444
        10.8%
        42%
        6.6
        HGV55BGXJ_n01_4QG3445
        12.7%
        42%
        3.8
        HGV55BGXJ_n01_4QG3447
        10.1%
        42%
        3.8
        HGV55BGXJ_n01_4QG3461
        11.3%
        42%
        4.8
        HGV55BGXJ_n01_4QG3475
        20.9%
        43%
        1.0
        HGV55BGXJ_n01_4QG3478
        13.6%
        42%
        13.0
        HGV55BGXJ_n01_4QG3482
        10.9%
        42%
        5.1
        HGV55BGXJ_n01_4QG3483
        12.4%
        42%
        4.7
        HGV55BGXJ_n01_4QG3484
        11.0%
        42%
        4.6
        HGV55BGXJ_n01_4QG3486
        9.9%
        42%
        3.1
        HGV55BGXJ_n01_4QG3487
        11.7%
        42%
        4.6
        HGV55BGXJ_n01_4QG3493
        10.2%
        42%
        3.8
        HGV55BGXJ_n01_4QG3495
        12.0%
        42%
        4.7
        HGV55BGXJ_n01_4QG3496
        11.7%
        42%
        8.1
        HGV55BGXJ_n01_4QG3502
        17.7%
        43%
        3.8
        HGV55BGXJ_n01_4QG3507
        13.2%
        42%
        2.6
        HGV55BGXJ_n01_4QG3509
        14.1%
        42%
        3.3
        HGV55BGXJ_n01_4QG3512
        11.3%
        42%
        4.1
        HGV55BGXJ_n01_undetermined
        68.4%
        43%
        15.8
        HGV55BGXJ_n02_4QG3117
        9.5%
        44%
        5.0
        HGV55BGXJ_n02_4QG3120
        9.2%
        44%
        3.7
        HGV55BGXJ_n02_4QG3126
        12.7%
        45%
        4.8
        HGV55BGXJ_n02_4QG3129
        9.4%
        43%
        6.3
        HGV55BGXJ_n02_4QG3155
        10.5%
        44%
        3.9
        HGV55BGXJ_n02_4QG3161
        8.7%
        43%
        3.7
        HGV55BGXJ_n02_4QG3172
        9.4%
        43%
        5.6
        HGV55BGXJ_n02_4QG3189
        10.7%
        43%
        6.9
        HGV55BGXJ_n02_4QG3197
        8.2%
        43%
        6.6
        HGV55BGXJ_n02_4QG3205
        10.9%
        43%
        6.3
        HGV55BGXJ_n02_4QG3209
        12.5%
        45%
        4.2
        HGV55BGXJ_n02_4QG3225
        9.6%
        44%
        6.0
        HGV55BGXJ_n02_4QG3229
        8.9%
        43%
        6.4
        HGV55BGXJ_n02_4QG3232
        8.3%
        43%
        5.8
        HGV55BGXJ_n02_4QG3248
        7.2%
        42%
        3.4
        HGV55BGXJ_n02_4QG3254
        10.3%
        43%
        5.9
        HGV55BGXJ_n02_4QG3257
        10.4%
        44%
        6.0
        HGV55BGXJ_n02_4QG3259
        9.6%
        44%
        2.6
        HGV55BGXJ_n02_4QG3262
        11.6%
        45%
        4.2
        HGV55BGXJ_n02_4QG3263
        11.4%
        45%
        4.4
        HGV55BGXJ_n02_4QG3264
        10.4%
        45%
        1.7
        HGV55BGXJ_n02_4QG3267
        8.8%
        44%
        2.1
        HGV55BGXJ_n02_4QG3270
        10.7%
        45%
        2.6
        HGV55BGXJ_n02_4QG3272
        8.5%
        43%
        2.5
        HGV55BGXJ_n02_4QG3273
        10.9%
        44%
        4.4
        HGV55BGXJ_n02_4QG3274
        16.5%
        47%
        2.0
        HGV55BGXJ_n02_4QG3289
        14.4%
        46%
        6.1
        HGV55BGXJ_n02_4QG3292
        10.2%
        44%
        5.6
        HGV55BGXJ_n02_4QG3297
        10.5%
        45%
        1.8
        HGV55BGXJ_n02_4QG3300
        8.9%
        43%
        5.1
        HGV55BGXJ_n02_4QG3301
        11.4%
        44%
        5.1
        HGV55BGXJ_n02_4QG3303
        9.0%
        44%
        3.6
        HGV55BGXJ_n02_4QG3307
        15.0%
        47%
        1.6
        HGV55BGXJ_n02_4QG3316
        10.3%
        44%
        2.6
        HGV55BGXJ_n02_4QG3320
        11.6%
        46%
        5.2
        HGV55BGXJ_n02_4QG3327
        20.5%
        49%
        3.0
        HGV55BGXJ_n02_4QG3332
        14.1%
        45%
        1.9
        HGV55BGXJ_n02_4QG3339
        14.6%
        46%
        4.4
        HGV55BGXJ_n02_4QG3345
        10.1%
        43%
        3.7
        HGV55BGXJ_n02_4QG3346
        12.1%
        44%
        5.1
        HGV55BGXJ_n02_4QG3349
        11.0%
        45%
        4.4
        HGV55BGXJ_n02_4QG3350
        12.7%
        44%
        7.7
        HGV55BGXJ_n02_4QG3352
        12.7%
        45%
        4.9
        HGV55BGXJ_n02_4QG3354
        11.0%
        44%
        4.9
        HGV55BGXJ_n02_4QG3355
        10.9%
        45%
        4.5
        HGV55BGXJ_n02_4QG3356
        14.6%
        46%
        2.7
        HGV55BGXJ_n02_4QG3357
        11.6%
        45%
        3.4
        HGV55BGXJ_n02_4QG3358
        12.3%
        46%
        1.3
        HGV55BGXJ_n02_4QG3360
        14.0%
        45%
        7.4
        HGV55BGXJ_n02_4QG3363
        11.2%
        45%
        2.5
        HGV55BGXJ_n02_4QG3365
        13.1%
        45%
        4.7
        HGV55BGXJ_n02_4QG3366
        13.8%
        46%
        4.4
        HGV55BGXJ_n02_4QG3367
        12.8%
        46%
        1.7
        HGV55BGXJ_n02_4QG3370
        12.5%
        46%
        2.2
        HGV55BGXJ_n02_4QG3372
        13.5%
        45%
        5.3
        HGV55BGXJ_n02_4QG3375
        10.8%
        45%
        1.7
        HGV55BGXJ_n02_4QG3387
        12.2%
        44%
        5.6
        HGV55BGXJ_n02_4QG3395
        15.0%
        46%
        2.4
        HGV55BGXJ_n02_4QG3400
        11.7%
        44%
        5.6
        HGV55BGXJ_n02_4QG3401
        12.6%
        45%
        3.7
        HGV55BGXJ_n02_4QG3404
        11.9%
        45%
        4.6
        HGV55BGXJ_n02_4QG3405
        12.3%
        46%
        3.7
        HGV55BGXJ_n02_4QG3407
        12.4%
        45%
        4.5
        HGV55BGXJ_n02_4QG3409
        10.4%
        45%
        3.8
        HGV55BGXJ_n02_4QG3412
        19.6%
        51%
        0.0
        HGV55BGXJ_n02_4QG3415
        9.2%
        44%
        3.6
        HGV55BGXJ_n02_4QG3421
        8.4%
        44%
        2.6
        HGV55BGXJ_n02_4QG3422
        11.2%
        44%
        4.9
        HGV55BGXJ_n02_4QG3425
        11.6%
        45%
        3.4
        HGV55BGXJ_n02_4QG3426
        10.1%
        45%
        3.1
        HGV55BGXJ_n02_4QG3430
        9.7%
        44%
        5.5
        HGV55BGXJ_n02_4QG3432
        9.5%
        44%
        4.2
        HGV55BGXJ_n02_4QG3436
        10.9%
        44%
        4.7
        HGV55BGXJ_n02_4QG3437
        11.5%
        45%
        3.3
        HGV55BGXJ_n02_4QG3441
        9.3%
        44%
        3.2
        HGV55BGXJ_n02_4QG3442
        14.5%
        47%
        2.6
        HGV55BGXJ_n02_4QG3444
        10.1%
        44%
        6.6
        HGV55BGXJ_n02_4QG3445
        11.5%
        45%
        3.8
        HGV55BGXJ_n02_4QG3447
        9.3%
        44%
        3.8
        HGV55BGXJ_n02_4QG3461
        10.4%
        44%
        4.8
        HGV55BGXJ_n02_4QG3475
        18.3%
        49%
        1.0
        HGV55BGXJ_n02_4QG3478
        12.8%
        44%
        13.0
        HGV55BGXJ_n02_4QG3482
        10.0%
        44%
        5.1
        HGV55BGXJ_n02_4QG3483
        11.4%
        45%
        4.7
        HGV55BGXJ_n02_4QG3484
        10.0%
        44%
        4.6
        HGV55BGXJ_n02_4QG3486
        9.1%
        44%
        3.1
        HGV55BGXJ_n02_4QG3487
        10.7%
        45%
        4.6
        HGV55BGXJ_n02_4QG3493
        9.4%
        44%
        3.8
        HGV55BGXJ_n02_4QG3495
        11.0%
        44%
        4.7
        HGV55BGXJ_n02_4QG3496
        11.0%
        44%
        8.1
        HGV55BGXJ_n02_4QG3502
        16.0%
        47%
        3.8
        HGV55BGXJ_n02_4QG3507
        12.0%
        45%
        2.6
        HGV55BGXJ_n02_4QG3509
        12.6%
        45%
        3.3
        HGV55BGXJ_n02_4QG3512
        10.2%
        44%
        4.1
        HGV55BGXJ_n02_undetermined
        67.7%
        45%
        15.8

        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 95/95 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        15790289
        3.8
        4QG3117
        4969506
        1.2
        4QG3197
        6619749
        1.6
        4QG3257
        6031155
        1.5
        4QG3273
        4375801
        1.1
        4QG3307
        1630743
        0.4
        4QG3349
        4433772
        1.1
        4QG3363
        2520782
        0.6
        4QG3395
        2400489
        0.6
        4QG3421
        2627895
        0.6
        4QG3441
        3164453
        0.8
        4QG3482
        5084850
        1.2
        4QG3120
        3721685
        0.9
        4QG3205
        6345922
        1.5
        4QG3259
        2635720
        0.6
        4QG3274
        2026927
        0.5
        4QG3316
        2581814
        0.6
        4QG3352
        4867726
        1.2
        4QG3365
        4693204
        1.1
        4QG3400
        5643176
        1.4
        4QG3422
        4888845
        1.2
        4QG3442
        2585560
        0.6
        4QG3483
        4737772
        1.1
        4QG3502
        3785191
        0.9
        4QG3126
        4774749
        1.2
        4QG3209
        4229891
        1.0
        4QG3262
        4206862
        1.0
        4QG3289
        6133859
        1.5
        4QG3320
        5224442
        1.3
        4QG3354
        4851419
        1.2
        4QG3366
        4384866
        1.1
        4QG3401
        3709768
        0.9
        4QG3425
        3407606
        0.8
        4QG3444
        6585435
        1.6
        4QG3484
        4597344
        1.1
        4QG3507
        2631246
        0.6
        4QG3129
        6314343
        1.5
        4QG3225
        6030431
        1.5
        4QG3263
        4391097
        1.1
        4QG3292
        5599666
        1.4
        4QG3327
        2964057
        0.7
        4QG3355
        4457716
        1.1
        4QG3367
        1715802
        0.4
        4QG3404
        4627415
        1.1
        4QG3426
        3062743
        0.7
        4QG3445
        3833360
        0.9
        4QG3486
        3083173
        0.7
        4QG3509
        3338621
        0.8
        4QG3155
        3942367
        1.0
        4QG3229
        6382281
        1.5
        4QG3264
        1698791
        0.4
        4QG3297
        1847885
        0.4
        4QG3332
        1872430
        0.5
        4QG3356
        2672692
        0.6
        4QG3370
        2216919
        0.5
        4QG3407
        4540170
        1.1
        4QG3430
        5469968
        1.3
        4QG3447
        3765170
        0.9
        4QG3487
        4589371
        1.1
        4QG3512
        4060279
        1.0
        4QG3161
        3697850
        0.9
        4QG3232
        5793869
        1.4
        4QG3267
        2124404
        0.5
        4QG3300
        5140320
        1.2
        4QG3339
        4395791
        1.1
        4QG3357
        3351628
        0.8
        4QG3372
        5279093
        1.3
        4QG3409
        3796406
        0.9
        4QG3432
        4216215
        1.0
        4QG3461
        4841693
        1.2
        4QG3493
        3836946
        0.9
        4QG3405
        3731931
        0.9
        4QG3172
        5634308
        1.4
        4QG3248
        3377215
        0.8
        4QG3270
        2625487
        0.6
        4QG3301
        5107931
        1.2
        4QG3345
        3655942
        0.9
        4QG3358
        1331394
        0.3
        4QG3375
        1684102
        0.4
        4QG3412
        44801
        0.0
        4QG3436
        4681152
        1.1
        4QG3475
        1016285
        0.2
        4QG3495
        4725158
        1.1
        4QG3350
        7713931
        1.9
        4QG3189
        6850715
        1.7
        4QG3254
        5883302
        1.4
        4QG3272
        2453531
        0.6
        4QG3303
        3552889
        0.9
        4QG3346
        5139000
        1.2
        4QG3360
        7362433
        1.8
        4QG3387
        5555722
        1.3
        4QG3415
        3630796
        0.9
        4QG3437
        3329807
        0.8
        4QG3478
        13015022
        3.2
        4QG3496
        8067052
        2.0

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        4.0
        439408808
        412021381
        3.8
        2.5

        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
        11587629.0
        73.4
        NNNNNNNN
        33382.0
        0.2
        GGGGGGGC
        26056.0
        0.2
        GGGGGGGT
        19829.0
        0.1
        GAACGTTA
        12858.0
        0.1
        GGGGGGTG
        12496.0
        0.1
        GGGGGGCC
        11346.0
        0.1
        GTGGGGGG
        10040.0
        0.1
        GGGGGGCG
        9448.0
        0.1
        CTCGACAA
        9395.0
        0.1
        AACGGTTA
        8698.0
        0.1
        TCGACTTA
        8447.0
        0.1
        CTTCGTTA
        8304.0
        0.1
        GGGGGCGG
        8088.0
        0.1
        GCGGGGGG
        7361.0
        0.1
        GGGGGCCC
        7311.0
        0.1
        TGCGTACA
        7265.0
        0.1
        GAAGGTTA
        7061.0
        0.0
        GGGGGGTT
        6941.0
        0.0
        TGCGAGAA
        6839.0
        0.0

        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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