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        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 2024-07-31, 16:06 based on data in: /vast/gencore/GENEFLOW/work/e2/4bc2a2283f43d51a23c1f1f9f38f9f/merged


        General Statistics

        Showing 98/98 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HKVNYDSXC_n01_CG_49
        33.1%
        41%
        308.2
        HKVNYDSXC_n01_CG_50
        32.7%
        39%
        220.0
        HKVNYDSXC_n01_CG_51
        41.0%
        40%
        549.7
        HKVNYDSXC_n01_CG_52
        32.9%
        39%
        271.0
        HKVNYDSXC_n01_CG_53
        37.0%
        39%
        440.9
        HKVNYDSXC_n01_CG_54
        38.3%
        39%
        481.3
        HKVNYDSXC_n01_CG_55
        35.9%
        42%
        600.8
        HKVNYDSXC_n01_CG_56
        36.7%
        39%
        363.3
        HKVNYDSXC_n01_CG_57
        43.2%
        40%
        815.0
        HKVNYDSXC_n01_CG_58
        40.7%
        40%
        523.8
        HKVNYDSXC_n01_CG_59
        33.8%
        40%
        148.1
        HKVNYDSXC_n01_CG_60
        43.3%
        40%
        580.7
        HKVNYDSXC_n01_CG_61
        32.0%
        40%
        129.6
        HKVNYDSXC_n01_CG_62
        37.0%
        40%
        323.5
        HKVNYDSXC_n01_CG_63
        42.8%
        40%
        677.1
        HKVNYDSXC_n01_CG_64
        40.1%
        40%
        448.2
        HKVNYDSXC_n01_CG_65
        33.1%
        41%
        139.6
        HKVNYDSXC_n01_CG_66
        31.6%
        40%
        120.5
        HKVNYDSXC_n01_CG_67
        29.6%
        40%
        154.5
        HKVNYDSXC_n01_CG_68
        26.2%
        40%
        75.2
        HKVNYDSXC_n01_CG_69
        33.5%
        40%
        219.8
        HKVNYDSXC_n01_CG_70
        45.8%
        39%
        963.2
        HKVNYDSXC_n01_CG_71
        39.4%
        39%
        496.4
        HKVNYDSXC_n01_CG_72
        30.0%
        40%
        115.6
        HKVNYDSXC_n01_CG_73
        26.4%
        39%
        72.8
        HKVNYDSXC_n01_CG_74
        37.5%
        39%
        415.4
        HKVNYDSXC_n01_CG_75
        30.0%
        40%
        164.5
        HKVNYDSXC_n01_CG_76
        32.3%
        39%
        140.2
        HKVNYDSXC_n01_CG_77
        30.6%
        40%
        98.5
        HKVNYDSXC_n01_CG_78
        31.8%
        39%
        153.4
        HKVNYDSXC_n01_CG_79
        35.6%
        39%
        253.1
        HKVNYDSXC_n01_CG_80
        34.3%
        40%
        143.6
        HKVNYDSXC_n01_CG_81
        29.0%
        40%
        65.1
        HKVNYDSXC_n01_CG_82
        30.1%
        40%
        74.3
        HKVNYDSXC_n01_CG_83
        32.2%
        39%
        124.5
        HKVNYDSXC_n01_CG_84
        29.3%
        40%
        72.2
        HKVNYDSXC_n01_CG_85
        24.9%
        40%
        34.3
        HKVNYDSXC_n01_CG_86
        30.3%
        39%
        112.5
        HKVNYDSXC_n01_CG_87
        24.0%
        44%
        18.5
        HKVNYDSXC_n01_CG_88
        33.6%
        39%
        145.1
        HKVNYDSXC_n01_CG_89
        27.3%
        40%
        55.7
        HKVNYDSXC_n01_CG_90
        30.7%
        39%
        84.8
        HKVNYDSXC_n01_CG_91
        31.3%
        39%
        154.2
        HKVNYDSXC_n01_CG_92
        28.4%
        39%
        85.9
        HKVNYDSXC_n01_CG_93
        25.7%
        40%
        47.0
        HKVNYDSXC_n01_CG_94
        27.1%
        39%
        66.3
        HKVNYDSXC_n01_CG_95
        28.9%
        39%
        92.3
        HKVNYDSXC_n01_CG_96
        27.8%
        40%
        73.3
        HKVNYDSXC_n01_undetermined
        68.5%
        41%
        936.7
        HKVNYDSXC_n02_CG_49
        32.5%
        41%
        308.2
        HKVNYDSXC_n02_CG_50
        32.0%
        39%
        220.0
        HKVNYDSXC_n02_CG_51
        40.1%
        40%
        549.7
        HKVNYDSXC_n02_CG_52
        32.4%
        39%
        271.0
        HKVNYDSXC_n02_CG_53
        36.2%
        39%
        440.9
        HKVNYDSXC_n02_CG_54
        37.7%
        39%
        481.3
        HKVNYDSXC_n02_CG_55
        34.9%
        42%
        600.8
        HKVNYDSXC_n02_CG_56
        35.8%
        39%
        363.3
        HKVNYDSXC_n02_CG_57
        41.9%
        39%
        815.0
        HKVNYDSXC_n02_CG_58
        39.6%
        40%
        523.8
        HKVNYDSXC_n02_CG_59
        32.7%
        40%
        148.1
        HKVNYDSXC_n02_CG_60
        42.3%
        40%
        580.7
        HKVNYDSXC_n02_CG_61
        31.1%
        40%
        129.6
        HKVNYDSXC_n02_CG_62
        36.1%
        40%
        323.5
        HKVNYDSXC_n02_CG_63
        42.0%
        40%
        677.1
        HKVNYDSXC_n02_CG_64
        39.3%
        40%
        448.2
        HKVNYDSXC_n02_CG_65
        32.7%
        41%
        139.6
        HKVNYDSXC_n02_CG_66
        31.0%
        40%
        120.5
        HKVNYDSXC_n02_CG_67
        29.1%
        40%
        154.5
        HKVNYDSXC_n02_CG_68
        25.7%
        40%
        75.2
        HKVNYDSXC_n02_CG_69
        32.9%
        40%
        219.8
        HKVNYDSXC_n02_CG_70
        44.7%
        39%
        963.2
        HKVNYDSXC_n02_CG_71
        38.4%
        39%
        496.4
        HKVNYDSXC_n02_CG_72
        29.3%
        40%
        115.6
        HKVNYDSXC_n02_CG_73
        25.8%
        39%
        72.8
        HKVNYDSXC_n02_CG_74
        36.1%
        39%
        415.4
        HKVNYDSXC_n02_CG_75
        29.4%
        40%
        164.5
        HKVNYDSXC_n02_CG_76
        31.3%
        39%
        140.2
        HKVNYDSXC_n02_CG_77
        30.1%
        40%
        98.5
        HKVNYDSXC_n02_CG_78
        31.1%
        39%
        153.4
        HKVNYDSXC_n02_CG_79
        35.0%
        39%
        253.1
        HKVNYDSXC_n02_CG_80
        33.7%
        40%
        143.6
        HKVNYDSXC_n02_CG_81
        28.4%
        40%
        65.1
        HKVNYDSXC_n02_CG_82
        29.3%
        40%
        74.3
        HKVNYDSXC_n02_CG_83
        31.8%
        39%
        124.5
        HKVNYDSXC_n02_CG_84
        28.8%
        40%
        72.2
        HKVNYDSXC_n02_CG_85
        24.3%
        40%
        34.3
        HKVNYDSXC_n02_CG_86
        29.6%
        39%
        112.5
        HKVNYDSXC_n02_CG_87
        23.2%
        43%
        18.5
        HKVNYDSXC_n02_CG_88
        33.1%
        39%
        145.1
        HKVNYDSXC_n02_CG_89
        26.8%
        40%
        55.7
        HKVNYDSXC_n02_CG_90
        30.1%
        39%
        84.8
        HKVNYDSXC_n02_CG_91
        30.8%
        39%
        154.2
        HKVNYDSXC_n02_CG_92
        28.1%
        39%
        85.9
        HKVNYDSXC_n02_CG_93
        25.2%
        40%
        47.0
        HKVNYDSXC_n02_CG_94
        26.5%
        39%
        66.3
        HKVNYDSXC_n02_CG_95
        28.3%
        39%
        92.3
        HKVNYDSXC_n02_CG_96
        27.5%
        40%
        73.3
        HKVNYDSXC_n02_undetermined
        67.6%
        42%
        936.7

        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 49/49 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        936684143
        7.3
        CG_49
        308169726
        2.4
        CG_50
        220034127
        1.7
        CG_51
        549673744
        4.3
        CG_52
        270969285
        2.1
        CG_53
        440935591
        3.4
        CG_54
        481299903
        3.7
        CG_55
        600811055
        4.7
        CG_56
        363307484
        2.8
        CG_57
        815013760
        6.3
        CG_58
        523789338
        4.1
        CG_59
        148136358
        1.2
        CG_60
        580747495
        4.5
        CG_61
        129571373
        1.0
        CG_62
        323502250
        2.5
        CG_63
        677081598
        5.3
        CG_64
        448230249
        3.5
        CG_65
        139629937
        1.1
        CG_66
        120530018
        0.9
        CG_67
        154489793
        1.2
        CG_68
        75176388
        0.6
        CG_69
        219796549
        1.7
        CG_70
        963206483
        7.5
        CG_71
        496351242
        3.9
        CG_72
        115584595
        0.9
        CG_73
        72757548
        0.6
        CG_74
        415436533
        3.2
        CG_75
        164486410
        1.3
        CG_76
        140156087
        1.1
        CG_77
        98527314
        0.8
        CG_78
        153430674
        1.2
        CG_79
        253114854
        2.0
        CG_80
        143575681
        1.1
        CG_81
        65124382
        0.5
        CG_82
        74297238
        0.6
        CG_83
        124546072
        1.0
        CG_84
        72202373
        0.6
        CG_85
        34282523
        0.3
        CG_86
        112464922
        0.9
        CG_87
        18500256
        0.1
        CG_88
        145053985
        1.1
        CG_89
        55693625
        0.4
        CG_90
        84769056
        0.7
        CG_91
        154249027
        1.2
        CG_92
        85942205
        0.7
        CG_93
        47046609
        0.4
        CG_94
        66280912
        0.5
        CG_95
        92298777
        0.7
        CG_96
        73299525
        0.6

        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. If your libraries are dual indexed, the two indices are concatenated.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGGGGAGATCTCG
        524344440.0
        56.0
        GGGGGGGGCTTCTGAG
        3961710.0
        0.4
        GGGGGGGGAGGTCACT
        3602983.0
        0.4
        GGGGGGGGCAAGGTCT
        2859230.0
        0.3
        GGGGGGGGGCAATTCG
        2843396.0
        0.3
        GGGGGGGGACGGTCTT
        2273992.0
        0.2
        GGGGGGGGGATAGGCT
        2194945.0
        0.2
        GGCAAGTTGGGGGGGG
        1711983.0
        0.2
        GGGGGGGGGATACTGG
        1647121.0
        0.2
        GGGGGGGGCTTAGGAC
        1532220.0
        0.2
        GGGGGGGGTGAACCTG
        1469024.0
        0.2
        GGGGGGGGACCTGGAA
        1383595.0
        0.1
        ATAACGCCGGGGGGGG
        1336548.0
        0.1
        GGGGGGGGAGCTCCTA
        1324631.0
        0.1
        GGGGGGGGCTGGAGTA
        1299846.0
        0.1
        GGGGGGGGCTTCGTTC
        1241623.0
        0.1
        TCTAGAGAAGGTCACT
        1110964.0
        0.1
        GCAAGTTACTTCTGAG
        1087251.0
        0.1
        AGCGAGATGGGGGGGG
        1020504.0
        0.1
        AGTGCTTAACCTGGAA
        1016933.0
        0.1

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        4.0
        15320088576
        12850259072
        7.3
        4.1

        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.

        loading..

        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.

        loading..

        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.

        loading..

        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.

        loading..

        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.

        loading..

        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.

        loading..

        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.

        loading..

        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.

        loading..

        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.

        loading..