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        Download the raw data used to create the plots in this report below:

        Note that additional data was saved in multiqc_data when this report was generated.


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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 2024-11-27, 15:35 based on data in: /scratch/gencore/GENEFLOW/work/nf/75/1727282bb952a636dd80e0aa8a142c/1


        General Statistics

        Showing 100/100 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH1
        45.4%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH10
        61.0%
        59%
        0.1
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH11
        31.2%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH12
        37.4%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH13
        37.2%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH14
        26.2%
        60%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH15
        37.2%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH16
        4.6%
        48%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH17
        30.6%
        62%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH18
        28.8%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH19
        29.7%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH2
        13.7%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH20
        27.9%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH21
        14.2%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH22
        19.7%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH23
        52.7%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH24
        30.0%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH25
        33.3%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH26
        26.9%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH27
        29.1%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH28
        27.8%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH29
        24.2%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH3
        32.3%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH30
        15.5%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH31
        48.4%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH32
        34.4%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH33
        42.7%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH34
        30.8%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH35
        32.0%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH36
        23.6%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH37
        31.7%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH38
        42.2%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH39
        43.9%
        62%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH4
        29.9%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH40
        35.7%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH41
        40.4%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH42
        29.7%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH43
        32.8%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH44
        26.5%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH45
        37.2%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH46
        26.7%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH47
        31.4%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH48
        37.6%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH49
        15.8%
        55%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH5
        37.4%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH50
        35.4%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH51
        32.1%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH52
        44.3%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH53
        42.0%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH54
        35.2%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH55
        46.7%
        64%
        0.1
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH56
        36.2%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH57
        29.6%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH58
        34.5%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH59
        25.8%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH6
        44.7%
        62%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH60
        49.8%
        62%
        0.1
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH61
        45.2%
        64%
        0.1
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH62
        34.5%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH63
        41.0%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH64
        27.9%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH65
        38.0%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH66
        33.3%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH67
        36.1%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH68
        39.7%
        61%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH69
        36.4%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH7
        30.5%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH70
        30.7%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH71
        35.5%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH72
        33.9%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH73
        39.6%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH74
        15.8%
        59%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH75
        39.8%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH76
        40.8%
        62%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH77
        22.1%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH78
        23.1%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH79
        29.9%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH8
        20.5%
        58%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH80
        33.7%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH81
        27.6%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH82
        21.8%
        62%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH83
        27.0%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH84
        29.6%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH85
        43.4%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH86
        23.2%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH87
        30.6%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH872
        43.7%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH873
        32.6%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH874
        34.5%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH88
        34.7%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH89
        43.0%
        64%
        0.1
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH9
        33.3%
        61%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH90
        33.4%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH91
        29.5%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH92
        29.0%
        63%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH93
        40.1%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH94
        26.2%
        64%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH95
        28.7%
        65%
        0.0
        000000000-GNC2B_l01_n01_micro_run2_repeat_GyrBATH96
        32.5%
        63%
        0.0
        000000000-GNC2B_l01_n01_undetermined
        84.8%
        46%
        0.8

        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 100/100 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        micro_run2_repeat_GyrBATH1
        40039
        1.0
        micro_run2_repeat_GyrBATH10
        61136
        1.6
        micro_run2_repeat_GyrBATH11
        35369
        0.9
        micro_run2_repeat_GyrBATH12
        37436
        1.0
        micro_run2_repeat_GyrBATH13
        47290
        1.2
        micro_run2_repeat_GyrBATH14
        22966
        0.6
        micro_run2_repeat_GyrBATH15
        44806
        1.2
        micro_run2_repeat_GyrBATH16
        7588
        0.2
        micro_run2_repeat_GyrBATH17
        18633
        0.5
        micro_run2_repeat_GyrBATH18
        35364
        0.9
        micro_run2_repeat_GyrBATH19
        28726
        0.8
        micro_run2_repeat_GyrBATH2
        665.0
        0.0
        micro_run2_repeat_GyrBATH20
        30004
        0.8
        micro_run2_repeat_GyrBATH21
        16139
        0.4
        micro_run2_repeat_GyrBATH22
        21349
        0.6
        micro_run2_repeat_GyrBATH23
        48344
        1.3
        micro_run2_repeat_GyrBATH24
        29077
        0.8
        micro_run2_repeat_GyrBATH25
        20045
        0.5
        micro_run2_repeat_GyrBATH26
        31266
        0.8
        micro_run2_repeat_GyrBATH27
        20454
        0.5
        micro_run2_repeat_GyrBATH28
        27050
        0.7
        micro_run2_repeat_GyrBATH29
        21890
        0.6
        micro_run2_repeat_GyrBATH3
        32905
        0.9
        micro_run2_repeat_GyrBATH30
        16278
        0.4
        micro_run2_repeat_GyrBATH31
        34961
        0.9
        micro_run2_repeat_GyrBATH32
        29714
        0.8
        micro_run2_repeat_GyrBATH33
        29297
        0.8
        micro_run2_repeat_GyrBATH34
        26679
        0.7
        micro_run2_repeat_GyrBATH35
        30497
        0.8
        micro_run2_repeat_GyrBATH36
        23698
        0.6
        micro_run2_repeat_GyrBATH37
        28698
        0.8
        micro_run2_repeat_GyrBATH38
        32386
        0.8
        micro_run2_repeat_GyrBATH39
        36703
        1.0
        micro_run2_repeat_GyrBATH4
        31875
        0.8
        micro_run2_repeat_GyrBATH40
        28630
        0.7
        micro_run2_repeat_GyrBATH41
        35406
        0.9
        micro_run2_repeat_GyrBATH42
        19621
        0.5
        micro_run2_repeat_GyrBATH43
        32869
        0.9
        micro_run2_repeat_GyrBATH44
        21969
        0.6
        micro_run2_repeat_GyrBATH45
        27683
        0.7
        micro_run2_repeat_GyrBATH46
        27362
        0.7
        micro_run2_repeat_GyrBATH47
        24188
        0.6
        micro_run2_repeat_GyrBATH48
        28803
        0.8
        micro_run2_repeat_GyrBATH49
        17195
        0.5
        micro_run2_repeat_GyrBATH5
        35837
        0.9
        micro_run2_repeat_GyrBATH50
        36122
        0.9
        micro_run2_repeat_GyrBATH51
        35781
        0.9
        micro_run2_repeat_GyrBATH52
        35892
        0.9
        micro_run2_repeat_GyrBATH53
        34961
        0.9
        micro_run2_repeat_GyrBATH54
        40436
        1.1
        micro_run2_repeat_GyrBATH55
        52002
        1.4
        micro_run2_repeat_GyrBATH56
        47236
        1.2
        micro_run2_repeat_GyrBATH57
        33294
        0.9
        micro_run2_repeat_GyrBATH58
        32158
        0.8
        micro_run2_repeat_GyrBATH59
        33187
        0.9
        micro_run2_repeat_GyrBATH6
        32581
        0.9
        micro_run2_repeat_GyrBATH60
        51218
        1.3
        micro_run2_repeat_GyrBATH61
        51173
        1.3
        micro_run2_repeat_GyrBATH62
        33085
        0.9
        micro_run2_repeat_GyrBATH63
        49891
        1.3
        micro_run2_repeat_GyrBATH64
        36385
        1.0
        micro_run2_repeat_GyrBATH65
        29137
        0.8
        micro_run2_repeat_GyrBATH66
        28459
        0.7
        micro_run2_repeat_GyrBATH67
        30932
        0.8
        micro_run2_repeat_GyrBATH68
        28920
        0.8
        micro_run2_repeat_GyrBATH69
        30318
        0.8
        micro_run2_repeat_GyrBATH7
        14214
        0.4
        micro_run2_repeat_GyrBATH70
        29107
        0.8
        micro_run2_repeat_GyrBATH71
        26242
        0.7
        micro_run2_repeat_GyrBATH72
        28708
        0.8
        micro_run2_repeat_GyrBATH73
        34088
        0.9
        micro_run2_repeat_GyrBATH74
        6536
        0.2
        micro_run2_repeat_GyrBATH75
        35539
        0.9
        micro_run2_repeat_GyrBATH76
        38569
        1.0
        micro_run2_repeat_GyrBATH77
        22706
        0.6
        micro_run2_repeat_GyrBATH78
        23129
        0.6
        micro_run2_repeat_GyrBATH79
        37977
        1.0
        micro_run2_repeat_GyrBATH8
        11386
        0.3
        micro_run2_repeat_GyrBATH80
        30900
        0.8
        micro_run2_repeat_GyrBATH81
        19898
        0.5
        micro_run2_repeat_GyrBATH82
        16542
        0.4
        micro_run2_repeat_GyrBATH83
        28488
        0.7
        micro_run2_repeat_GyrBATH84
        31634
        0.8
        micro_run2_repeat_GyrBATH85
        35485
        0.9
        micro_run2_repeat_GyrBATH86
        25425
        0.7
        micro_run2_repeat_GyrBATH87
        39830
        1.0
        micro_run2_repeat_GyrBATH872
        45344
        1.2
        micro_run2_repeat_GyrBATH873
        28269
        0.7
        micro_run2_repeat_GyrBATH874
        43656
        1.1
        micro_run2_repeat_GyrBATH88
        36867
        1.0
        micro_run2_repeat_GyrBATH89
        59514
        1.6
        micro_run2_repeat_GyrBATH9
        23070
        0.6
        micro_run2_repeat_GyrBATH90
        28879
        0.8
        micro_run2_repeat_GyrBATH91
        28273
        0.7
        micro_run2_repeat_GyrBATH92
        24405
        0.6
        micro_run2_repeat_GyrBATH93
        27415
        0.7
        micro_run2_repeat_GyrBATH94
        22660
        0.6
        micro_run2_repeat_GyrBATH95
        35957
        0.9
        micro_run2_repeat_GyrBATH96
        29222
        0.8
        undetermined
        772104
        20.2

        Barcodes of Undetermined Reads


        We have determined the barcodes of your undetermined reads. Here are the top 20 barcodes. The full list is available here.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        TCTAGACT-TCTTTCCC
        5425.0
        0.7
        TCAGTCTA-TCTTTCCC
        5393.0
        0.7
        AGCATACC-TCTTTCCC
        5342.0
        0.7
        ATAGCGCT-TCTTTCCC
        4404.0
        0.6
        CATCGTGA-TCTTTCCC
        4371.0
        0.6
        CGTCATAC-TCTTTCCC
        4268.0
        0.6
        TCCTCATG-TCTTTCCC
        4138.0
        0.5
        ATGAGCTC-TCTTTCCC
        4051.0
        0.5
        CGAGCTAG-TCTTTCCC
        3282.0
        0.4
        CTCTAGAG-TCTTTCCC
        3093.0
        0.4
        GAGCTCGA-TCTTTCCC
        3018.0
        0.4
        CTCTCTCT-TCTTTCCC
        2920.0
        0.4
        CAGTAGGT-TCTTTCCC
        2831.0
        0.4
        TCCTCATT-TCTTTCCC
        2705.0
        0.3
        CTCGACTT-TCTTTCCC
        2311.0
        0.3
        TCATTCTA-TCTTTCCC
        1861.0
        0.2
        TCTATACT-TCTTTCCC
        1728.0
        0.2
        TCCTCCTT-TCTTTCCC
        1616.0
        0.2
        TTTTTTTT-TCTTTCCC
        1600.0
        0.2
        ATCATACC-TCTTTCCC
        1484.0
        0.2

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        LaneTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        1.0
        4475826
        3824096
        20.2
        17.5

        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.

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        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.

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        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.

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        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.

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        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.

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        Sequence Length Distribution

        All samples have sequences of a single length (301bp).

        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.

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        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.

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        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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