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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 2022-02-25, 11:10 based on data in: /scratch/gencore/logs/html/HVGTVDRXY/merged


        General Statistics

        Showing 100/100 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HVGTVDRXY_n01_WT1_C_11
        6.2%
        40%
        0.6
        HVGTVDRXY_n01_WT1_C_2
        24.9%
        37%
        33.3
        HVGTVDRXY_n01_WT1_C_24
        22.2%
        37%
        16.4
        HVGTVDRXY_n01_WT1_C_48
        23.3%
        39%
        19.0
        HVGTVDRXY_n01_WT1_C_5
        7.4%
        42%
        23.7
        HVGTVDRXY_n01_WT1_C_72
        52.5%
        40%
        30.7
        HVGTVDRXY_n01_WT1_C_8
        12.1%
        41%
        32.9
        HVGTVDRXY_n01_WT2_C_11
        13.1%
        42%
        33.3
        HVGTVDRXY_n01_WT2_C_2
        12.6%
        37%
        30.4
        HVGTVDRXY_n01_WT2_C_24
        22.3%
        36%
        23.3
        HVGTVDRXY_n01_WT2_C_48
        16.8%
        41%
        24.8
        HVGTVDRXY_n01_WT2_C_5
        8.5%
        41%
        33.9
        HVGTVDRXY_n01_WT2_C_72
        40.8%
        40%
        22.1
        HVGTVDRXY_n01_WT2_C_8
        11.3%
        43%
        33.7
        HVGTVDRXY_n01_WT2_N_11
        13.3%
        36%
        0.7
        HVGTVDRXY_n01_WT2_N_2
        24.4%
        35%
        10.1
        HVGTVDRXY_n01_WT2_N_24
        39.2%
        37%
        7.4
        HVGTVDRXY_n01_WT2_N_48
        40.9%
        37%
        4.3
        HVGTVDRXY_n01_WT2_N_5
        26.3%
        35%
        7.5
        HVGTVDRXY_n01_WT2_N_72
        35.9%
        39%
        11.3
        HVGTVDRXY_n01_WT2_N_8
        23.7%
        37%
        1.8
        HVGTVDRXY_n01_WT2_Y_11
        10.5%
        39%
        23.4
        HVGTVDRXY_n01_WT2_Y_2
        9.8%
        39%
        34.7
        HVGTVDRXY_n01_WT2_Y_24
        13.5%
        38%
        23.5
        HVGTVDRXY_n01_WT2_Y_48
        20.4%
        37%
        11.9
        HVGTVDRXY_n01_WT2_Y_5
        7.7%
        44%
        31.7
        HVGTVDRXY_n01_WT2_Y_72
        24.6%
        48%
        3.8
        HVGTVDRXY_n01_WT2_Y_8
        9.3%
        44%
        30.8
        HVGTVDRXY_n01_WT3_C_11
        8.9%
        44%
        27.8
        HVGTVDRXY_n01_WT3_C_2
        15.0%
        36%
        25.7
        HVGTVDRXY_n01_WT3_C_24
        16.5%
        42%
        45.1
        HVGTVDRXY_n01_WT3_C_48
        24.7%
        39%
        41.6
        HVGTVDRXY_n01_WT3_C_5
        9.2%
        43%
        40.2
        HVGTVDRXY_n01_WT3_C_72
        42.8%
        37%
        41.6
        HVGTVDRXY_n01_WT3_C_8
        8.4%
        44%
        33.1
        HVGTVDRXY_n01_WT3_N_11
        29.5%
        35%
        19.7
        HVGTVDRXY_n01_WT3_N_2
        13.7%
        38%
        26.6
        HVGTVDRXY_n01_WT3_N_24
        27.6%
        36%
        13.0
        HVGTVDRXY_n01_WT3_N_48
        37.3%
        36%
        6.8
        HVGTVDRXY_n01_WT3_N_5
        25.9%
        35%
        17.1
        HVGTVDRXY_n01_WT3_N_72
        26.0%
        36%
        8.6
        HVGTVDRXY_n01_WT3_N_8
        27.2%
        35%
        12.5
        HVGTVDRXY_n01_WT3_Y_11
        23.1%
        38%
        16.5
        HVGTVDRXY_n01_WT3_Y_2
        12.4%
        38%
        30.2
        HVGTVDRXY_n01_WT3_Y_24
        13.3%
        37%
        15.0
        HVGTVDRXY_n01_WT3_Y_48
        11.6%
        39%
        24.2
        HVGTVDRXY_n01_WT3_Y_5
        10.2%
        44%
        41.6
        HVGTVDRXY_n01_WT3_Y_72
        26.2%
        36%
        5.1
        HVGTVDRXY_n01_WT3_Y_8
        7.4%
        41%
        21.5
        HVGTVDRXY_n01_undetermined
        59.7%
        42%
        35.8
        HVGTVDRXY_n02_WT1_C_11
        18.3%
        40%
        0.6
        HVGTVDRXY_n02_WT1_C_2
        44.2%
        36%
        33.3
        HVGTVDRXY_n02_WT1_C_24
        40.5%
        35%
        16.4
        HVGTVDRXY_n02_WT1_C_48
        44.9%
        39%
        19.0
        HVGTVDRXY_n02_WT1_C_5
        44.3%
        42%
        23.7
        HVGTVDRXY_n02_WT1_C_72
        62.4%
        40%
        30.7
        HVGTVDRXY_n02_WT1_C_8
        48.0%
        41%
        32.9
        HVGTVDRXY_n02_WT2_C_11
        54.4%
        42%
        33.3
        HVGTVDRXY_n02_WT2_C_2
        36.8%
        36%
        30.4
        HVGTVDRXY_n02_WT2_C_24
        40.3%
        35%
        23.3
        HVGTVDRXY_n02_WT2_C_48
        52.5%
        41%
        24.8
        HVGTVDRXY_n02_WT2_C_5
        44.8%
        41%
        33.9
        HVGTVDRXY_n02_WT2_C_72
        54.0%
        40%
        22.1
        HVGTVDRXY_n02_WT2_C_8
        56.7%
        42%
        33.7
        HVGTVDRXY_n02_WT2_N_11
        15.8%
        35%
        0.7
        HVGTVDRXY_n02_WT2_N_2
        32.9%
        33%
        10.1
        HVGTVDRXY_n02_WT2_N_24
        42.9%
        36%
        7.4
        HVGTVDRXY_n02_WT2_N_48
        41.5%
        36%
        4.3
        HVGTVDRXY_n02_WT2_N_5
        31.7%
        35%
        7.5
        HVGTVDRXY_n02_WT2_N_72
        39.0%
        38%
        11.3
        HVGTVDRXY_n02_WT2_N_8
        26.5%
        37%
        1.8
        HVGTVDRXY_n02_WT2_Y_11
        34.6%
        38%
        23.4
        HVGTVDRXY_n02_WT2_Y_2
        39.3%
        39%
        34.7
        HVGTVDRXY_n02_WT2_Y_24
        34.7%
        37%
        23.5
        HVGTVDRXY_n02_WT2_Y_48
        28.1%
        37%
        11.9
        HVGTVDRXY_n02_WT2_Y_5
        49.3%
        44%
        31.7
        HVGTVDRXY_n02_WT2_Y_72
        42.5%
        47%
        3.8
        HVGTVDRXY_n02_WT2_Y_8
        59.7%
        44%
        30.8
        HVGTVDRXY_n02_WT3_C_11
        54.5%
        44%
        27.8
        HVGTVDRXY_n02_WT3_C_2
        37.3%
        34%
        25.7
        HVGTVDRXY_n02_WT3_C_24
        56.1%
        42%
        45.1
        HVGTVDRXY_n02_WT3_C_48
        45.6%
        39%
        41.6
        HVGTVDRXY_n02_WT3_C_5
        48.3%
        43%
        40.2
        HVGTVDRXY_n02_WT3_C_72
        48.2%
        37%
        41.6
        HVGTVDRXY_n02_WT3_C_8
        50.8%
        44%
        33.1
        HVGTVDRXY_n02_WT3_N_11
        38.3%
        34%
        19.7
        HVGTVDRXY_n02_WT3_N_2
        40.8%
        36%
        26.6
        HVGTVDRXY_n02_WT3_N_24
        34.9%
        35%
        13.0
        HVGTVDRXY_n02_WT3_N_48
        39.1%
        36%
        6.8
        HVGTVDRXY_n02_WT3_N_5
        39.7%
        34%
        17.1
        HVGTVDRXY_n02_WT3_N_72
        30.2%
        36%
        8.6
        HVGTVDRXY_n02_WT3_N_8
        34.5%
        34%
        12.5
        HVGTVDRXY_n02_WT3_Y_11
        38.2%
        35%
        16.5
        HVGTVDRXY_n02_WT3_Y_2
        36.9%
        37%
        30.2
        HVGTVDRXY_n02_WT3_Y_24
        28.6%
        36%
        15.0
        HVGTVDRXY_n02_WT3_Y_48
        38.7%
        39%
        24.2
        HVGTVDRXY_n02_WT3_Y_5
        60.6%
        44%
        41.6
        HVGTVDRXY_n02_WT3_Y_72
        32.2%
        34%
        5.1
        HVGTVDRXY_n02_WT3_Y_8
        41.6%
        41%
        21.5
        HVGTVDRXY_n02_undetermined
        63.8%
        42%
        35.8

        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 50/50 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        35768546
        3.2
        WT1_C_2
        33290564
        3.0
        WT1_C_5
        23664955
        2.1
        WT1_C_8
        32856562
        3.0
        WT1_C_11
        563293
        0.1
        WT1_C_24
        16370665
        1.5
        WT1_C_48
        19030658
        1.7
        WT1_C_72
        30663061
        2.8
        WT2_C_2
        30408186
        2.7
        WT2_C_5
        33875032
        3.1
        WT2_C_8
        33719823
        3.0
        WT2_C_11
        33270607
        3.0
        WT2_C_24
        23257897
        2.1
        WT2_C_48
        24765561
        2.2
        WT2_C_72
        22068443
        2.0
        WT3_C_2
        25741637
        2.3
        WT3_C_5
        40166824
        3.6
        WT3_C_8
        33068496
        3.0
        WT3_C_11
        27778043
        2.5
        WT3_C_24
        45110548
        4.1
        WT3_C_48
        41605830
        3.7
        WT3_C_72
        41647675
        3.8
        WT2_Y_2
        34737366
        3.1
        WT2_Y_5
        31724092
        2.9
        WT2_Y_8
        30755762
        2.8
        WT2_Y_11
        23398243
        2.1
        WT2_Y_24
        23549573
        2.1
        WT2_Y_48
        11904859
        1.1
        WT2_Y_72
        3761587
        0.3
        WT3_Y_2
        30231940
        2.7
        WT3_Y_5
        41550216
        3.7
        WT3_Y_8
        21477249
        1.9
        WT3_Y_11
        16475739
        1.5
        WT3_Y_24
        14970491
        1.3
        WT3_Y_48
        24158226
        2.2
        WT3_Y_72
        5083577
        0.5
        WT2_N_2
        10147421
        0.9
        WT2_N_5
        7544881
        0.7
        WT2_N_8
        1757310
        0.2
        WT2_N_11
        727115
        0.1
        WT2_N_24
        7430626
        0.7
        WT2_N_48
        4320499
        0.4
        WT2_N_72
        11335970
        1.0
        WT3_N_2
        26625414
        2.4
        WT3_N_5
        17120709
        1.5
        WT3_N_8
        12536256
        1.1
        WT3_N_11
        19689699
        1.8
        WT3_N_24
        13046119
        1.2
        WT3_N_48
        6770636
        0.6
        WT3_N_72
        8604373
        0.8

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        2.0
        1276674048
        1110128854
        3.2
        1.8

        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 (%)
        GGGGGG
        26445794.0
        73.9
        TGATTA
        143317.0
        0.4
        CTAAGA
        129630.0
        0.4
        TTATGA
        124360.0
        0.3
        TAACTA
        109085.0
        0.3
        TAGTCA
        107059.0
        0.3
        TGGTAA
        104702.0
        0.3
        AAAAAA
        103974.0
        0.3
        TTGTAA
        89163.0
        0.2
        GGGGGC
        86720.0
        0.2
        GGATTA
        76491.0
        0.2
        CGTTAA
        69964.0
        0.2
        TACTTA
        63376.0
        0.2
        AGCGTA
        60582.0
        0.2
        GAGGTA
        60527.0
        0.2
        GGTTAA
        59684.0
        0.2
        AACAAA
        54985.0
        0.1
        TAGGAA
        54466.0
        0.1
        ATACGA
        54425.0
        0.1
        ATGTCA
        53804.0
        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.

        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 (101bp).

        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..