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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 2022-08-16, 23:35 based on data in: /scratch/gencore/logs/html/HKCGLBGXM/merged


        General Statistics

        Showing 194/194 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HKCGLBGXM_n01_1315
        22.4%
        41%
        1.0
        HKCGLBGXM_n01_1657
        19.7%
        41%
        0.8
        HKCGLBGXM_n01_2835
        18.8%
        43%
        0.9
        HKCGLBGXM_n01_2836
        20.6%
        43%
        1.0
        HKCGLBGXM_n01_2837
        20.5%
        42%
        0.9
        HKCGLBGXM_n01_2838
        18.3%
        42%
        0.7
        HKCGLBGXM_n01_2839
        18.3%
        43%
        0.8
        HKCGLBGXM_n01_2840
        17.4%
        41%
        0.5
        HKCGLBGXM_n01_2841
        18.5%
        42%
        1.0
        HKCGLBGXM_n01_2842
        19.3%
        42%
        1.1
        HKCGLBGXM_n01_2843
        15.3%
        42%
        0.6
        HKCGLBGXM_n01_2844
        18.6%
        43%
        0.9
        HKCGLBGXM_n01_2845
        16.5%
        41%
        0.6
        HKCGLBGXM_n01_2846
        17.6%
        41%
        0.9
        HKCGLBGXM_n01_2847
        16.9%
        41%
        0.7
        HKCGLBGXM_n01_2848
        17.9%
        41%
        0.9
        HKCGLBGXM_n01_2849
        16.2%
        41%
        0.6
        HKCGLBGXM_n01_2850
        17.4%
        42%
        0.7
        HKCGLBGXM_n01_2851
        17.8%
        40%
        0.4
        HKCGLBGXM_n01_2852
        19.7%
        42%
        1.0
        HKCGLBGXM_n01_2853
        18.1%
        42%
        1.0
        HKCGLBGXM_n01_2854
        15.5%
        42%
        0.6
        HKCGLBGXM_n01_2855
        15.7%
        42%
        0.7
        HKCGLBGXM_n01_2856
        17.6%
        41%
        0.9
        HKCGLBGXM_n01_2878
        14.6%
        41%
        0.6
        HKCGLBGXM_n01_2879
        15.9%
        40%
        0.5
        HKCGLBGXM_n01_2880
        14.2%
        40%
        0.5
        HKCGLBGXM_n01_2881
        22.3%
        42%
        0.7
        HKCGLBGXM_n01_2882
        16.2%
        40%
        0.5
        HKCGLBGXM_n01_2883
        18.6%
        39%
        0.3
        HKCGLBGXM_n01_2884
        19.7%
        40%
        0.5
        HKCGLBGXM_n01_2885
        18.2%
        41%
        0.7
        HKCGLBGXM_n01_2886
        13.2%
        40%
        0.3
        HKCGLBGXM_n01_2887
        13.4%
        40%
        0.4
        HKCGLBGXM_n01_2888
        13.4%
        39%
        0.4
        HKCGLBGXM_n01_2889
        16.5%
        40%
        0.6
        HKCGLBGXM_n01_2890
        20.4%
        41%
        0.7
        HKCGLBGXM_n01_2891
        18.0%
        41%
        0.8
        HKCGLBGXM_n01_2892
        15.8%
        40%
        0.6
        HKCGLBGXM_n01_2893
        13.9%
        40%
        0.4
        HKCGLBGXM_n01_2894
        14.0%
        40%
        0.4
        HKCGLBGXM_n01_2895
        18.9%
        40%
        0.5
        HKCGLBGXM_n01_2896
        17.7%
        41%
        0.8
        HKCGLBGXM_n01_2897
        23.0%
        40%
        0.6
        HKCGLBGXM_n01_2898
        10.0%
        40%
        0.3
        HKCGLBGXM_n01_2899
        15.7%
        40%
        0.5
        HKCGLBGXM_n01_2900
        17.2%
        40%
        0.7
        HKCGLBGXM_n01_2901
        18.3%
        40%
        0.8
        HKCGLBGXM_n01_2902
        17.8%
        41%
        0.8
        HKCGLBGXM_n01_2903
        18.9%
        41%
        0.8
        HKCGLBGXM_n01_2904
        15.6%
        40%
        0.5
        HKCGLBGXM_n01_2905
        14.9%
        41%
        0.3
        HKCGLBGXM_n01_2906
        16.3%
        40%
        0.4
        HKCGLBGXM_n01_2907
        23.1%
        41%
        0.7
        HKCGLBGXM_n01_2908
        19.0%
        40%
        0.5
        HKCGLBGXM_n01_2909
        21.1%
        40%
        0.4
        HKCGLBGXM_n01_2910
        21.6%
        40%
        0.3
        HKCGLBGXM_n01_2911
        15.4%
        40%
        0.6
        HKCGLBGXM_n01_2912
        18.2%
        40%
        0.3
        HKCGLBGXM_n01_2913
        18.0%
        40%
        0.6
        HKCGLBGXM_n01_2914
        20.0%
        42%
        0.8
        HKCGLBGXM_n01_2915
        17.9%
        42%
        0.6
        HKCGLBGXM_n01_2916
        16.9%
        41%
        0.6
        HKCGLBGXM_n01_2917
        16.1%
        41%
        0.5
        HKCGLBGXM_n01_2918
        17.7%
        42%
        0.8
        HKCGLBGXM_n01_2919
        16.6%
        42%
        0.6
        HKCGLBGXM_n01_2920
        20.2%
        41%
        0.5
        HKCGLBGXM_n01_2921
        16.7%
        41%
        0.5
        HKCGLBGXM_n01_2922
        18.5%
        40%
        0.3
        HKCGLBGXM_n01_2923
        17.6%
        41%
        0.7
        HKCGLBGXM_n01_2924
        14.2%
        40%
        0.4
        HKCGLBGXM_n01_2925
        14.8%
        40%
        0.5
        HKCGLBGXM_n01_2926
        14.2%
        40%
        0.4
        HKCGLBGXM_n01_2927
        13.5%
        40%
        0.5
        HKCGLBGXM_n01_2928
        13.2%
        40%
        0.3
        HKCGLBGXM_n01_2929
        11.4%
        40%
        0.3
        HKCGLBGXM_n01_2930
        15.3%
        40%
        0.4
        HKCGLBGXM_n01_2931
        10.3%
        40%
        0.1
        HKCGLBGXM_n01_2932
        11.4%
        40%
        0.3
        HKCGLBGXM_n01_2933
        14.0%
        40%
        0.4
        HKCGLBGXM_n01_2934
        15.9%
        41%
        0.7
        HKCGLBGXM_n01_2935
        18.9%
        41%
        0.9
        HKCGLBGXM_n01_2936
        16.4%
        41%
        0.8
        HKCGLBGXM_n01_2937
        14.4%
        40%
        0.5
        HKCGLBGXM_n01_2938
        15.5%
        41%
        0.6
        HKCGLBGXM_n01_2939
        15.7%
        40%
        0.4
        HKCGLBGXM_n01_2940
        17.7%
        40%
        0.5
        HKCGLBGXM_n01_2941
        16.5%
        41%
        0.6
        HKCGLBGXM_n01_2942
        9.4%
        39%
        0.2
        HKCGLBGXM_n01_2943
        12.4%
        40%
        0.4
        HKCGLBGXM_n01_2944
        17.8%
        40%
        0.6
        HKCGLBGXM_n01_2945
        15.8%
        41%
        0.5
        HKCGLBGXM_n01_2946
        18.2%
        40%
        0.7
        HKCGLBGXM_n01_2947
        16.2%
        40%
        0.4
        HKCGLBGXM_n01_2948
        15.1%
        40%
        0.6
        HKCGLBGXM_n01_DGY1
        19.9%
        43%
        0.8
        HKCGLBGXM_n01_undetermined
        90.6%
        44%
        19.5
        HKCGLBGXM_n02_1315
        22.4%
        41%
        1.0
        HKCGLBGXM_n02_1657
        19.8%
        41%
        0.8
        HKCGLBGXM_n02_2835
        19.1%
        42%
        0.9
        HKCGLBGXM_n02_2836
        20.7%
        43%
        1.0
        HKCGLBGXM_n02_2837
        20.7%
        42%
        0.9
        HKCGLBGXM_n02_2838
        18.4%
        42%
        0.7
        HKCGLBGXM_n02_2839
        18.5%
        42%
        0.8
        HKCGLBGXM_n02_2840
        17.5%
        40%
        0.5
        HKCGLBGXM_n02_2841
        18.7%
        42%
        1.0
        HKCGLBGXM_n02_2842
        19.6%
        42%
        1.1
        HKCGLBGXM_n02_2843
        15.4%
        42%
        0.6
        HKCGLBGXM_n02_2844
        18.7%
        42%
        0.9
        HKCGLBGXM_n02_2845
        16.7%
        41%
        0.6
        HKCGLBGXM_n02_2846
        17.7%
        41%
        0.9
        HKCGLBGXM_n02_2847
        16.9%
        41%
        0.7
        HKCGLBGXM_n02_2848
        18.1%
        41%
        0.9
        HKCGLBGXM_n02_2849
        16.3%
        41%
        0.6
        HKCGLBGXM_n02_2850
        17.7%
        41%
        0.7
        HKCGLBGXM_n02_2851
        17.9%
        40%
        0.4
        HKCGLBGXM_n02_2852
        20.0%
        41%
        1.0
        HKCGLBGXM_n02_2853
        18.2%
        41%
        1.0
        HKCGLBGXM_n02_2854
        15.5%
        42%
        0.6
        HKCGLBGXM_n02_2855
        16.0%
        41%
        0.7
        HKCGLBGXM_n02_2856
        17.7%
        41%
        0.9
        HKCGLBGXM_n02_2878
        14.9%
        41%
        0.6
        HKCGLBGXM_n02_2879
        16.1%
        40%
        0.5
        HKCGLBGXM_n02_2880
        14.4%
        40%
        0.5
        HKCGLBGXM_n02_2881
        22.4%
        41%
        0.7
        HKCGLBGXM_n02_2882
        16.4%
        40%
        0.5
        HKCGLBGXM_n02_2883
        18.7%
        39%
        0.3
        HKCGLBGXM_n02_2884
        19.9%
        40%
        0.5
        HKCGLBGXM_n02_2885
        18.3%
        40%
        0.7
        HKCGLBGXM_n02_2886
        13.2%
        40%
        0.3
        HKCGLBGXM_n02_2887
        13.6%
        39%
        0.4
        HKCGLBGXM_n02_2888
        13.6%
        39%
        0.4
        HKCGLBGXM_n02_2889
        16.6%
        40%
        0.6
        HKCGLBGXM_n02_2890
        20.6%
        41%
        0.7
        HKCGLBGXM_n02_2891
        18.2%
        41%
        0.8
        HKCGLBGXM_n02_2892
        16.0%
        40%
        0.6
        HKCGLBGXM_n02_2893
        14.1%
        40%
        0.4
        HKCGLBGXM_n02_2894
        14.3%
        40%
        0.4
        HKCGLBGXM_n02_2895
        19.1%
        40%
        0.5
        HKCGLBGXM_n02_2896
        17.9%
        40%
        0.8
        HKCGLBGXM_n02_2897
        23.2%
        40%
        0.6
        HKCGLBGXM_n02_2898
        10.3%
        39%
        0.3
        HKCGLBGXM_n02_2899
        15.8%
        40%
        0.5
        HKCGLBGXM_n02_2900
        17.3%
        40%
        0.7
        HKCGLBGXM_n02_2901
        18.6%
        40%
        0.8
        HKCGLBGXM_n02_2902
        18.0%
        41%
        0.8
        HKCGLBGXM_n02_2903
        18.8%
        41%
        0.8
        HKCGLBGXM_n02_2904
        15.8%
        40%
        0.5
        HKCGLBGXM_n02_2905
        14.8%
        41%
        0.3
        HKCGLBGXM_n02_2906
        16.5%
        40%
        0.4
        HKCGLBGXM_n02_2907
        23.2%
        41%
        0.7
        HKCGLBGXM_n02_2908
        19.1%
        40%
        0.5
        HKCGLBGXM_n02_2909
        21.2%
        40%
        0.4
        HKCGLBGXM_n02_2910
        21.7%
        40%
        0.3
        HKCGLBGXM_n02_2911
        15.6%
        40%
        0.6
        HKCGLBGXM_n02_2912
        18.3%
        40%
        0.3
        HKCGLBGXM_n02_2913
        18.0%
        40%
        0.6
        HKCGLBGXM_n02_2914
        20.1%
        42%
        0.8
        HKCGLBGXM_n02_2915
        17.9%
        42%
        0.6
        HKCGLBGXM_n02_2916
        17.0%
        41%
        0.6
        HKCGLBGXM_n02_2917
        16.3%
        40%
        0.5
        HKCGLBGXM_n02_2918
        17.8%
        41%
        0.8
        HKCGLBGXM_n02_2919
        16.7%
        41%
        0.6
        HKCGLBGXM_n02_2920
        20.1%
        41%
        0.5
        HKCGLBGXM_n02_2921
        16.7%
        41%
        0.5
        HKCGLBGXM_n02_2922
        18.7%
        40%
        0.3
        HKCGLBGXM_n02_2923
        17.7%
        41%
        0.7
        HKCGLBGXM_n02_2924
        14.4%
        40%
        0.4
        HKCGLBGXM_n02_2925
        15.0%
        40%
        0.5
        HKCGLBGXM_n02_2926
        14.2%
        40%
        0.4
        HKCGLBGXM_n02_2927
        13.4%
        40%
        0.5
        HKCGLBGXM_n02_2928
        13.3%
        40%
        0.3
        HKCGLBGXM_n02_2929
        11.6%
        40%
        0.3
        HKCGLBGXM_n02_2930
        15.5%
        40%
        0.4
        HKCGLBGXM_n02_2931
        10.4%
        40%
        0.1
        HKCGLBGXM_n02_2932
        11.5%
        39%
        0.3
        HKCGLBGXM_n02_2933
        14.2%
        40%
        0.4
        HKCGLBGXM_n02_2934
        16.0%
        41%
        0.7
        HKCGLBGXM_n02_2935
        18.9%
        41%
        0.9
        HKCGLBGXM_n02_2936
        16.6%
        41%
        0.8
        HKCGLBGXM_n02_2937
        14.6%
        40%
        0.5
        HKCGLBGXM_n02_2938
        15.9%
        40%
        0.6
        HKCGLBGXM_n02_2939
        15.8%
        40%
        0.4
        HKCGLBGXM_n02_2940
        17.8%
        40%
        0.5
        HKCGLBGXM_n02_2941
        16.6%
        41%
        0.6
        HKCGLBGXM_n02_2942
        9.7%
        39%
        0.2
        HKCGLBGXM_n02_2943
        12.6%
        40%
        0.4
        HKCGLBGXM_n02_2944
        17.9%
        40%
        0.6
        HKCGLBGXM_n02_2945
        15.9%
        41%
        0.5
        HKCGLBGXM_n02_2946
        18.3%
        40%
        0.7
        HKCGLBGXM_n02_2947
        16.4%
        40%
        0.4
        HKCGLBGXM_n02_2948
        15.3%
        40%
        0.6
        HKCGLBGXM_n02_DGY1
        19.9%
        42%
        0.8
        HKCGLBGXM_n02_undetermined
        90.0%
        44%
        19.5

        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 97/97 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        19466871
        25.4
        2835.0
        869560
        1.1
        2846.0
        858239
        1.1
        2923.0
        712743
        0.9
        2934.0
        676574
        0.9
        2878.0
        573473
        0.7
        2890.0
        684086
        0.9
        2902.0
        796402
        1.0
        2914.0
        770902
        1.0
        2836.0
        1023739
        1.3
        2847.0
        660273
        0.9
        2924.0
        424415
        0.6
        2935.0
        864594
        1.1
        2879.0
        529588
        0.7
        2891.0
        784060
        1.0
        2903.0
        817712
        1.1
        2915.0
        629041
        0.8
        2837.0
        941230
        1.2
        2848.0
        925819
        1.2
        2925.0
        467767
        0.6
        2936.0
        771137
        1.0
        2880.0
        538102
        0.7
        2892.0
        637073
        0.8
        2904.0
        547310
        0.7
        2916.0
        572279
        0.7
        2838.0
        662103
        0.9
        2849.0
        620533
        0.8
        2926.0
        377226
        0.5
        2937.0
        538347
        0.7
        2881.0
        661701
        0.9
        2893.0
        389169
        0.5
        2905.0
        289651
        0.4
        2917.0
        521505
        0.7
        2839.0
        781345
        1.0
        2850.0
        701713
        0.9
        2927.0
        491556
        0.6
        2938.0
        605916
        0.8
        2882.0
        542624
        0.7
        2894.0
        360263
        0.5
        2906.0
        393452
        0.5
        2918.0
        784894
        1.0
        2840.0
        533159
        0.7
        2851.0
        405672
        0.5
        2928.0
        304787
        0.4
        2939.0
        426569
        0.6
        2883.0
        339589
        0.4
        2895.0
        538393
        0.7
        2907.0
        683713
        0.9
        2919.0
        585648
        0.8
        2841.0
        1008872
        1.3
        2852.0
        1039430
        1.4
        2929.0
        325797
        0.4
        2940.0
        509032
        0.7
        2884.0
        462527
        0.6
        2896.0
        830358
        1.1
        2908.0
        475865
        0.6
        2920.0
        520371
        0.7
        2842.0
        1146773
        1.5
        2853.0
        972386
        1.3
        2930.0
        350153
        0.5
        2941.0
        611198
        0.8
        2885.0
        669189
        0.9
        2897.0
        622637
        0.8
        2909.0
        432463
        0.6
        2921.0
        515457
        0.7
        2843.0
        550216
        0.7
        2854.0
        641551
        0.8
        2931.0
        139211
        0.2
        2942.0
        230808
        0.3
        2886.0
        267725
        0.3
        2898.0
        259760
        0.3
        2910.0
        345989
        0.5
        2922.0
        312967
        0.4
        2844.0
        896262
        1.2
        2855.0
        710661
        0.9
        2932.0
        263900
        0.3
        2943.0
        387190
        0.5
        2887.0
        379263
        0.5
        2899.0
        523373
        0.7
        2911.0
        551945
        0.7
        DGY1
        824136
        1.1
        2845.0
        630402
        0.8
        2856.0
        875630
        1.1
        2933.0
        371336
        0.5
        2944.0
        558949
        0.7
        2888.0
        369622
        0.5
        2900.0
        686166
        0.9
        2912.0
        345972
        0.5
        1315.0
        1011594
        1.3
        2945.0
        500555
        0.7
        2946.0
        650895
        0.8
        2947.0
        445263
        0.6
        2948.0
        615570
        0.8
        2889.0
        607842
        0.8
        2901.0
        835662
        1.1
        2913.0
        586891
        0.8
        1657.0
        827059
        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
        78922162
        76779390
        25.4
        22.1

        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 (%)
        GGGGGGGGAGATCTCG
        17063315.0
        87.7
        GGGGGGGGAGAGGATA
        102152.0
        0.5
        GGGGGGGGATAGAGAG
        91014.0
        0.5
        GGGGGGGGTACTCCTT
        66584.0
        0.3
        GGGGGGGGTCTTACGC
        64698.0
        0.3
        GGGGGGGGCTCCTTAC
        60118.0
        0.3
        GGGGGGGGAGGCTTAG
        60010.0
        0.3
        GGGGGGGGTATGCAGT
        56993.0
        0.3
        GGGGGGGGTCTACTCT
        42557.0
        0.2
        GCTAGCTAAGAGGATA
        12532.0
        0.1
        GCTAGCTAATAGAGAG
        12355.0
        0.1
        AGAGAGGAAGAGGATA
        8685.0
        0.0
        TAAGGCGAGGGGGGGG
        8636.0
        0.0
        GCTAGCTAAGGCTTAG
        8366.0
        0.0
        NNNNNNNNNNNNNNNN
        7676.0
        0.0
        GTAGAGGAGGGGGGGG
        7513.0
        0.0
        CGTACTAGGGGGGGGG
        7465.0
        0.0
        AGGCAGAAGGGGGGGG
        7295.0
        0.0
        GCTAGCTATATGCAGT
        7202.0
        0.0
        GCTAGCTATCTTACGC
        6883.0
        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 (151bp).

        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.

        194 samples had less than 1% of reads made up of overrepresented sequences

        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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