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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 2021-06-28, 16:50 based on data in: /scratch/gencore/logs/html/H53C5BGXJ/merged


        General Statistics

        Showing 192/192 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        H53C5BGXJ_n01_3QG3087
        6.7%
        41%
        2.6
        H53C5BGXJ_n01_3QG3098
        11.1%
        41%
        10.0
        H53C5BGXJ_n01_3QG3102
        10.9%
        42%
        3.8
        H53C5BGXJ_n01_3QG3108
        13.4%
        40%
        1.3
        H53C5BGXJ_n01_3QG3110
        11.4%
        42%
        3.2
        H53C5BGXJ_n01_3QG3114
        9.4%
        42%
        4.8
        H53C5BGXJ_n01_3QG3123
        9.1%
        42%
        5.4
        H53C5BGXJ_n01_3QG3131
        11.3%
        41%
        11.4
        H53C5BGXJ_n01_3QG3134
        5.3%
        41%
        0.7
        H53C5BGXJ_n01_3QG3137
        6.6%
        42%
        2.9
        H53C5BGXJ_n01_3QG3142
        10.1%
        42%
        3.0
        H53C5BGXJ_n01_3QG3151
        6.0%
        42%
        0.4
        H53C5BGXJ_n01_3QG3152
        9.5%
        42%
        7.3
        H53C5BGXJ_n01_3QG3157
        0.2%
        40%
        0.0
        H53C5BGXJ_n01_3QG3164
        8.2%
        42%
        3.7
        H53C5BGXJ_n01_3QG3166
        8.1%
        42%
        2.6
        H53C5BGXJ_n01_3QG3175
        10.1%
        42%
        4.7
        H53C5BGXJ_n01_3QG3178
        7.6%
        41%
        5.6
        H53C5BGXJ_n01_3QG3183
        5.9%
        42%
        1.7
        H53C5BGXJ_n01_3QG3187
        7.7%
        42%
        4.1
        H53C5BGXJ_n01_3QG3188
        7.2%
        42%
        5.1
        H53C5BGXJ_n01_3QG3191
        10.2%
        42%
        9.3
        H53C5BGXJ_n01_3QG3199
        7.2%
        42%
        0.9
        H53C5BGXJ_n01_3QG3202
        8.8%
        42%
        5.6
        H53C5BGXJ_n01_3QG3203
        8.5%
        42%
        4.1
        H53C5BGXJ_n01_3QG3206
        7.2%
        42%
        3.8
        H53C5BGXJ_n01_3QG3208
        10.1%
        42%
        5.1
        H53C5BGXJ_n01_3QG3217
        8.3%
        42%
        2.1
        H53C5BGXJ_n01_3QG3218
        9.5%
        41%
        8.3
        H53C5BGXJ_n01_3QG3223
        9.3%
        42%
        0.9
        H53C5BGXJ_n01_3QG3226
        9.6%
        42%
        6.5
        H53C5BGXJ_n01_3QG3230
        8.2%
        42%
        4.9
        H53C5BGXJ_n01_3QG3231
        9.5%
        42%
        8.3
        H53C5BGXJ_n01_3QG3233
        8.6%
        42%
        1.6
        H53C5BGXJ_n01_3QG3234
        7.8%
        41%
        3.2
        H53C5BGXJ_n01_3QG3235
        8.6%
        41%
        5.5
        H53C5BGXJ_n01_3QG3242
        9.1%
        42%
        0.7
        H53C5BGXJ_n01_3QG3245
        6.2%
        42%
        4.3
        H53C5BGXJ_n01_3QG3255
        10.3%
        42%
        5.4
        H53C5BGXJ_n01_3QG3260
        7.6%
        42%
        5.2
        H53C5BGXJ_n01_3QG3268
        9.6%
        41%
        4.3
        H53C5BGXJ_n01_3QG3281
        8.8%
        42%
        4.1
        H53C5BGXJ_n01_3QG3284
        9.6%
        42%
        9.8
        H53C5BGXJ_n01_3QG3287
        7.9%
        42%
        4.2
        H53C5BGXJ_n01_3QG3288
        8.5%
        42%
        0.9
        H53C5BGXJ_n01_3QG3290
        11.4%
        41%
        3.9
        H53C5BGXJ_n01_3QG3291
        10.2%
        42%
        4.9
        H53C5BGXJ_n01_3QG3293
        8.8%
        41%
        7.4
        H53C5BGXJ_n01_3QG3294
        7.9%
        42%
        2.7
        H53C5BGXJ_n01_3QG3296
        7.8%
        41%
        2.6
        H53C5BGXJ_n01_3QG3317
        10.8%
        43%
        10.7
        H53C5BGXJ_n01_3QG3325
        11.5%
        42%
        0.3
        H53C5BGXJ_n01_3QG3335
        9.1%
        41%
        7.6
        H53C5BGXJ_n01_3QG3337
        3.8%
        41%
        0.6
        H53C5BGXJ_n01_3QG3340
        9.2%
        42%
        7.3
        H53C5BGXJ_n01_3QG3342
        10.5%
        41%
        9.4
        H53C5BGXJ_n01_3QG3343
        10.5%
        41%
        10.3
        H53C5BGXJ_n01_3QG3344
        9.4%
        41%
        3.3
        H53C5BGXJ_n01_3QG3347
        12.7%
        41%
        9.6
        H53C5BGXJ_n01_3QG3351
        9.1%
        41%
        4.9
        H53C5BGXJ_n01_3QG3353
        11.2%
        41%
        10.8
        H53C5BGXJ_n01_3QG3364
        4.9%
        39%
        0.8
        H53C5BGXJ_n01_3QG3368
        7.2%
        41%
        3.3
        H53C5BGXJ_n01_3QG3376
        8.6%
        41%
        2.2
        H53C5BGXJ_n01_3QG3377
        8.6%
        41%
        2.7
        H53C5BGXJ_n01_3QG3381
        10.4%
        41%
        5.6
        H53C5BGXJ_n01_3QG3382
        9.5%
        42%
        5.1
        H53C5BGXJ_n01_3QG3383
        10.4%
        41%
        5.3
        H53C5BGXJ_n01_3QG3385
        11.1%
        41%
        6.5
        H53C5BGXJ_n01_3QG3388
        13.1%
        41%
        9.2
        H53C5BGXJ_n01_3QG3391
        7.6%
        40%
        0.4
        H53C5BGXJ_n01_3QG3392
        5.7%
        42%
        1.7
        H53C5BGXJ_n01_3QG3394
        11.4%
        42%
        1.1
        H53C5BGXJ_n01_3QG3397
        10.0%
        42%
        3.0
        H53C5BGXJ_n01_3QG3399
        9.6%
        42%
        4.7
        H53C5BGXJ_n01_3QG3403
        8.1%
        41%
        1.6
        H53C5BGXJ_n01_3QG3406
        12.3%
        41%
        9.5
        H53C5BGXJ_n01_3QG3420
        10.5%
        41%
        9.1
        H53C5BGXJ_n01_3QG3427
        10.6%
        42%
        10.6
        H53C5BGXJ_n01_3QG3428
        9.2%
        41%
        7.3
        H53C5BGXJ_n01_3QG3431
        8.5%
        42%
        5.4
        H53C5BGXJ_n01_3QG3433
        7.4%
        42%
        1.7
        H53C5BGXJ_n01_3QG3434
        8.2%
        41%
        2.7
        H53C5BGXJ_n01_3QG3438
        6.7%
        42%
        2.1
        H53C5BGXJ_n01_3QG3439
        9.1%
        42%
        5.2
        H53C5BGXJ_n01_3QG3440
        7.4%
        42%
        4.4
        H53C5BGXJ_n01_3QG3443
        3.9%
        41%
        0.4
        H53C5BGXJ_n01_3QG3446
        11.0%
        42%
        9.9
        H53C5BGXJ_n01_3QG3452
        10.5%
        42%
        9.3
        H53C5BGXJ_n01_3QG3454
        7.2%
        42%
        1.8
        H53C5BGXJ_n01_3QG3456
        11.0%
        42%
        10.0
        H53C5BGXJ_n01_3QG3458
        11.7%
        41%
        4.7
        H53C5BGXJ_n01_3QG3464
        8.6%
        42%
        1.5
        H53C5BGXJ_n01_3QG3468
        8.7%
        42%
        4.3
        H53C5BGXJ_n01_3QG3470
        11.7%
        42%
        1.9
        H53C5BGXJ_n01_undetermined
        67.7%
        43%
        17.3
        H53C5BGXJ_n02_3QG3087
        6.2%
        42%
        2.6
        H53C5BGXJ_n02_3QG3098
        10.5%
        42%
        10.0
        H53C5BGXJ_n02_3QG3102
        10.1%
        43%
        3.8
        H53C5BGXJ_n02_3QG3108
        11.6%
        43%
        1.3
        H53C5BGXJ_n02_3QG3110
        10.0%
        44%
        3.2
        H53C5BGXJ_n02_3QG3114
        8.8%
        42%
        4.8
        H53C5BGXJ_n02_3QG3123
        8.6%
        42%
        5.4
        H53C5BGXJ_n02_3QG3131
        10.6%
        42%
        11.4
        H53C5BGXJ_n02_3QG3134
        4.7%
        42%
        0.7
        H53C5BGXJ_n02_3QG3137
        6.1%
        43%
        2.9
        H53C5BGXJ_n02_3QG3142
        9.2%
        43%
        3.0
        H53C5BGXJ_n02_3QG3151
        5.2%
        43%
        0.4
        H53C5BGXJ_n02_3QG3152
        8.9%
        43%
        7.3
        H53C5BGXJ_n02_3QG3157
        6.5%
        44%
        0.0
        H53C5BGXJ_n02_3QG3164
        7.4%
        43%
        3.7
        H53C5BGXJ_n02_3QG3166
        7.3%
        43%
        2.6
        H53C5BGXJ_n02_3QG3175
        9.1%
        43%
        4.7
        H53C5BGXJ_n02_3QG3178
        7.1%
        42%
        5.6
        H53C5BGXJ_n02_3QG3183
        5.2%
        43%
        1.7
        H53C5BGXJ_n02_3QG3187
        7.0%
        43%
        4.1
        H53C5BGXJ_n02_3QG3188
        6.6%
        43%
        5.1
        H53C5BGXJ_n02_3QG3191
        9.4%
        42%
        9.3
        H53C5BGXJ_n02_3QG3199
        6.1%
        43%
        0.9
        H53C5BGXJ_n02_3QG3202
        8.2%
        43%
        5.6
        H53C5BGXJ_n02_3QG3203
        7.7%
        43%
        4.1
        H53C5BGXJ_n02_3QG3206
        6.6%
        43%
        3.8
        H53C5BGXJ_n02_3QG3208
        9.3%
        43%
        5.1
        H53C5BGXJ_n02_3QG3217
        7.1%
        44%
        2.1
        H53C5BGXJ_n02_3QG3218
        8.9%
        42%
        8.3
        H53C5BGXJ_n02_3QG3223
        8.1%
        44%
        0.9
        H53C5BGXJ_n02_3QG3226
        8.9%
        43%
        6.5
        H53C5BGXJ_n02_3QG3230
        7.6%
        42%
        4.9
        H53C5BGXJ_n02_3QG3231
        9.0%
        42%
        8.3
        H53C5BGXJ_n02_3QG3233
        7.5%
        44%
        1.6
        H53C5BGXJ_n02_3QG3234
        7.2%
        42%
        3.2
        H53C5BGXJ_n02_3QG3235
        8.2%
        42%
        5.5
        H53C5BGXJ_n02_3QG3242
        7.5%
        44%
        0.7
        H53C5BGXJ_n02_3QG3245
        5.8%
        42%
        4.3
        H53C5BGXJ_n02_3QG3255
        9.6%
        43%
        5.4
        H53C5BGXJ_n02_3QG3260
        7.2%
        43%
        5.2
        H53C5BGXJ_n02_3QG3268
        8.7%
        42%
        4.3
        H53C5BGXJ_n02_3QG3281
        7.9%
        43%
        4.1
        H53C5BGXJ_n02_3QG3284
        9.1%
        42%
        9.8
        H53C5BGXJ_n02_3QG3287
        7.3%
        43%
        4.2
        H53C5BGXJ_n02_3QG3288
        7.3%
        43%
        0.9
        H53C5BGXJ_n02_3QG3290
        10.2%
        43%
        3.9
        H53C5BGXJ_n02_3QG3291
        9.4%
        43%
        4.9
        H53C5BGXJ_n02_3QG3293
        8.4%
        42%
        7.4
        H53C5BGXJ_n02_3QG3294
        7.3%
        43%
        2.7
        H53C5BGXJ_n02_3QG3296
        7.2%
        42%
        2.6
        H53C5BGXJ_n02_3QG3317
        10.2%
        44%
        10.7
        H53C5BGXJ_n02_3QG3325
        10.2%
        46%
        0.3
        H53C5BGXJ_n02_3QG3335
        8.7%
        42%
        7.6
        H53C5BGXJ_n02_3QG3337
        3.4%
        41%
        0.6
        H53C5BGXJ_n02_3QG3340
        8.8%
        42%
        7.3
        H53C5BGXJ_n02_3QG3342
        9.8%
        42%
        9.4
        H53C5BGXJ_n02_3QG3343
        9.9%
        42%
        10.3
        H53C5BGXJ_n02_3QG3344
        8.5%
        42%
        3.3
        H53C5BGXJ_n02_3QG3347
        11.9%
        42%
        9.6
        H53C5BGXJ_n02_3QG3351
        8.7%
        42%
        4.9
        H53C5BGXJ_n02_3QG3353
        10.6%
        42%
        10.8
        H53C5BGXJ_n02_3QG3364
        4.5%
        40%
        0.8
        H53C5BGXJ_n02_3QG3368
        6.6%
        42%
        3.3
        H53C5BGXJ_n02_3QG3376
        7.7%
        42%
        2.2
        H53C5BGXJ_n02_3QG3377
        7.8%
        42%
        2.7
        H53C5BGXJ_n02_3QG3381
        9.6%
        42%
        5.6
        H53C5BGXJ_n02_3QG3382
        8.7%
        43%
        5.1
        H53C5BGXJ_n02_3QG3383
        9.6%
        42%
        5.3
        H53C5BGXJ_n02_3QG3385
        10.3%
        42%
        6.5
        H53C5BGXJ_n02_3QG3388
        12.5%
        42%
        9.2
        H53C5BGXJ_n02_3QG3391
        6.5%
        42%
        0.4
        H53C5BGXJ_n02_3QG3392
        5.0%
        43%
        1.7
        H53C5BGXJ_n02_3QG3394
        9.7%
        45%
        1.1
        H53C5BGXJ_n02_3QG3397
        8.9%
        43%
        3.0
        H53C5BGXJ_n02_3QG3399
        8.8%
        43%
        4.7
        H53C5BGXJ_n02_3QG3403
        7.0%
        43%
        1.6
        H53C5BGXJ_n02_3QG3406
        11.7%
        42%
        9.5
        H53C5BGXJ_n02_3QG3420
        9.8%
        42%
        9.1
        H53C5BGXJ_n02_3QG3427
        10.2%
        42%
        10.6
        H53C5BGXJ_n02_3QG3428
        8.8%
        42%
        7.3
        H53C5BGXJ_n02_3QG3431
        8.0%
        43%
        5.4
        H53C5BGXJ_n02_3QG3433
        6.5%
        43%
        1.7
        H53C5BGXJ_n02_3QG3434
        7.6%
        42%
        2.7
        H53C5BGXJ_n02_3QG3438
        6.1%
        43%
        2.1
        H53C5BGXJ_n02_3QG3439
        8.3%
        43%
        5.2
        H53C5BGXJ_n02_3QG3440
        6.9%
        42%
        4.4
        H53C5BGXJ_n02_3QG3443
        3.4%
        42%
        0.4
        H53C5BGXJ_n02_3QG3446
        10.4%
        42%
        9.9
        H53C5BGXJ_n02_3QG3452
        9.9%
        42%
        9.3
        H53C5BGXJ_n02_3QG3454
        6.4%
        43%
        1.8
        H53C5BGXJ_n02_3QG3456
        10.5%
        42%
        10.0
        H53C5BGXJ_n02_3QG3458
        11.0%
        42%
        4.7
        H53C5BGXJ_n02_3QG3464
        7.3%
        44%
        1.5
        H53C5BGXJ_n02_3QG3468
        8.1%
        42%
        4.3
        H53C5BGXJ_n02_3QG3470
        10.3%
        44%
        1.9
        H53C5BGXJ_n02_undetermined
        67.1%
        43%
        17.3

        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
        17289371
        3.7
        3QG3108
        1335176
        0.3
        3QG3391
        444626
        0.1
        3QG3364
        762214
        0.2
        3QG3293
        7443837
        1.6
        3QG3382
        5086228
        1.1
        3QG3468
        4319169
        0.9
        3QG3383
        5261822
        1.1
        3QG3098
        9991239
        2.2
        3QG3420
        9118167
        2.0
        3QG3385
        6490646
        1.4
        3QG3114
        4765981
        1.0
        3QG3294
        2689401
        0.6
        3QG3178
        5620937
        1.2
        3QG3137
        2910879
        0.6
        3QG3434
        2691021
        0.6
        3QG3377
        2719034
        0.6
        3QG3242
        739407
        0.2
        3QG3403
        1585099
        0.3
        3QG3175
        4707744
        1.0
        3QG3340
        7344338
        1.6
        3QG3470
        1881062
        0.4
        3QG3431
        5352806
        1.2
        3QG3164
        3716975
        0.8
        3QG3230
        4926219
        1.1
        3QG3284
        9804118
        2.1
        3QG3335
        7644178
        1.7
        3QG3087
        2649014
        0.6
        3QG3123
        5448859
        1.2
        3QG3183
        1660746
        0.4
        3QG3268
        4316893
        0.9
        3QG3131
        11431855
        2.5
        3QG3188
        5105117
        1.1
        3QG3231
        8345788
        1.8
        3QG3446
        9903176
        2.1
        3QG3287
        4234675
        0.9
        3QG3157
        572.0
        0.0
        3QG3317
        10723798
        2.3
        3QG3187
        4051572
        0.9
        3QG3368
        3349767
        0.7
        3QG3223
        901113
        0.2
        3QG3351
        4871453
        1.1
        3QG3342
        9369821
        2.0
        3QG3353
        10827420
        2.3
        3QG3202
        5589153
        1.2
        3QG3218
        8256438
        1.8
        3QG3102
        3838790
        0.8
        3QG3191
        9300787
        2.0
        3QG3110
        3230600
        0.7
        3QG3456
        10047761
        2.2
        3QG3381
        5600340
        1.2
        3QG3427
        10584662
        2.3
        3QG3296
        2640551
        0.6
        3QG3376
        2241031
        0.5
        3QG3290
        3943666
        0.9
        3QG3206
        3812217
        0.8
        3QG3452
        9341619
        2.0
        3QG3439
        5238492
        1.1
        3QG3152
        7260673
        1.6
        3QG3291
        4850821
        1.0
        3QG3281
        4080842
        0.9
        3QG3343
        10274463
        2.2
        3QG3428
        7307752
        1.6
        3QG3347
        9606505
        2.1
        3QG3388
        9249898
        2.0
        3QG3440
        4435656
        1.0
        3QG3344
        3289492
        0.7
        3QG3166
        2619346
        0.6
        3QG3208
        5082108
        1.1
        3QG3235
        5503970
        1.2
        3QG3226
        6519558
        1.4
        3QG3255
        5398116
        1.2
        3QG3217
        2114582
        0.5
        3QG3406
        9508291
        2.1
        3QG3245
        4335990
        0.9
        3QG3392
        1706352
        0.4
        3QG3234
        3175715
        0.7
        3QG3394
        1068513
        0.2
        3QG3433
        1731787
        0.4
        3QG3199
        949048
        0.2
        3QG3454
        1777109
        0.4
        3QG3134
        704505
        0.2
        3QG3458
        4683441
        1.0
        3QG3399
        4655238
        1.0
        3QG3397
        2965931
        0.6
        3QG3142
        3037189
        0.7
        3QG3337
        550811
        0.1
        3QG3443
        418360
        0.1
        3QG3151
        425911
        0.1
        3QG3288
        850669
        0.2
        3QG3233
        1638220
        0.4
        3QG3325
        347362
        0.1
        3QG3464
        1524510
        0.3
        3QG3438
        2072188
        0.4
        3QG3203
        4120289
        0.9
        3QG3260
        5229668
        1.1

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        4.0
        494689152
        462570319
        3.7
        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
        13027381.0
        75.3
        NNNNNNNN
        77749.0
        0.5
        GGGGGGGC
        25997.0
        0.1
        GGGGGGGT
        18633.0
        0.1
        NNNNNNNC
        15808.0
        0.1
        GGGGGGTG
        14859.0
        0.1
        GTGGGGGG
        14727.0
        0.1
        NNNNNNNA
        12848.0
        0.1
        NNNNNNNT
        12222.0
        0.1
        CAGGTCAA
        11849.0
        0.1
        TCCATGCA
        11714.0
        0.1
        GAACGTTA
        11679.0
        0.1
        CAGTTCAA
        11497.0
        0.1
        GCGGGGGG
        11326.0
        0.1
        CTCAGAGA
        11270.0
        0.1
        TCCAGGTA
        11180.0
        0.1
        CCAGTGAA
        10998.0
        0.1
        GGGGGGCG
        10798.0
        0.1
        AGACCGTA
        10784.0
        0.1
        CTACAGCA
        10516.0
        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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