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        If you use plots from MultiQC in a publication or presentation, please cite:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

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        About MultiQC

        This report was generated using MultiQC, version 1.9

        You can see a YouTube video describing how to use MultiQC reports here: https://youtu.be/qPbIlO_KWN0

        For more information about MultiQC, including other videos and extensive documentation, please visit http://multiqc.info

        You can report bugs, suggest improvements and find the source code for MultiQC on GitHub: https://github.com/ewels/MultiQC

        MultiQC is published in Bioinformatics:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

        A modular tool to aggregate results from bioinformatics analyses across many samples into a single report.

        Report generated on 2021-11-08, 19:46 based on data in: /scratch/gencore/logs/html/HJMY5DRXY/merged


        General Statistics

        Showing 78/78 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HJMY5DRXY_n01_dpr402
        63.8%
        52%
        33.0
        HJMY5DRXY_n01_dpr403
        63.0%
        52%
        35.5
        HJMY5DRXY_n01_dpr404
        67.0%
        52%
        53.9
        HJMY5DRXY_n01_dpr405
        58.2%
        52%
        30.5
        HJMY5DRXY_n01_dpr406
        63.6%
        51%
        32.3
        HJMY5DRXY_n01_dpr407
        68.3%
        53%
        60.9
        HJMY5DRXY_n01_dpr408
        60.7%
        52%
        33.2
        HJMY5DRXY_n01_dpr409
        61.8%
        52%
        29.5
        HJMY5DRXY_n01_dpr410
        87.8%
        52%
        713.6
        HJMY5DRXY_n01_dpr411
        61.9%
        52%
        36.4
        HJMY5DRXY_n01_dpr412
        60.2%
        52%
        28.5
        HJMY5DRXY_n01_dpr413
        59.2%
        52%
        32.4
        HJMY5DRXY_n01_dpr414
        62.9%
        52%
        25.9
        HJMY5DRXY_n01_dpr415
        66.7%
        54%
        36.7
        HJMY5DRXY_n01_dpr416
        74.0%
        55%
        34.7
        HJMY5DRXY_n01_dpr417
        66.8%
        54%
        37.1
        HJMY5DRXY_n01_dpr418
        73.8%
        54%
        37.9
        HJMY5DRXY_n01_dpr419
        73.2%
        55%
        40.6
        HJMY5DRXY_n01_dpr420
        72.8%
        54%
        41.0
        HJMY5DRXY_n01_dpr421
        67.8%
        53%
        34.8
        HJMY5DRXY_n01_dpr422
        74.8%
        55%
        35.2
        HJMY5DRXY_n01_dpr423
        63.4%
        52%
        40.9
        HJMY5DRXY_n01_dpr424
        63.5%
        51%
        36.5
        HJMY5DRXY_n01_dpr425
        60.6%
        51%
        36.8
        HJMY5DRXY_n01_dpr426
        60.0%
        51%
        26.0
        HJMY5DRXY_n01_dpr427
        57.7%
        51%
        30.4
        HJMY5DRXY_n01_dpr428
        56.6%
        51%
        28.3
        HJMY5DRXY_n01_dpr429
        56.9%
        51%
        28.6
        HJMY5DRXY_n01_dpr430
        62.5%
        51%
        26.6
        HJMY5DRXY_n01_dpr431
        58.5%
        51%
        29.9
        HJMY5DRXY_n01_dpr432
        58.3%
        51%
        28.5
        HJMY5DRXY_n01_dpr433
        55.7%
        51%
        24.2
        HJMY5DRXY_n01_dpr434
        62.9%
        52%
        28.1
        HJMY5DRXY_n01_dpr435
        60.7%
        52%
        32.0
        HJMY5DRXY_n01_dpr436
        60.6%
        52%
        28.6
        HJMY5DRXY_n01_dpr437
        68.6%
        55%
        29.7
        HJMY5DRXY_n01_dpr438
        46.4%
        50%
        1.5
        HJMY5DRXY_n01_dpr439
        30.6%
        49%
        0.4
        HJMY5DRXY_n01_dpr440
        39.8%
        49%
        0.6
        HJMY5DRXY_n01_dpr441
        28.8%
        50%
        0.5
        HJMY5DRXY_n01_dpr442
        46.9%
        51%
        4.6
        HJMY5DRXY_n01_dpr443
        36.9%
        51%
        3.3
        HJMY5DRXY_n01_dpr444
        51.9%
        51%
        3.5
        HJMY5DRXY_n01_dpr445
        0.1%
        50%
        0.0
        HJMY5DRXY_n01_dpr446
        4.2%
        50%
        0.0
        HJMY5DRXY_n01_dpr447
        32.4%
        51%
        2.4
        HJMY5DRXY_n01_dpr448
        25.7%
        51%
        1.5
        HJMY5DRXY_n01_dpr449
        33.7%
        51%
        1.8
        HJMY5DRXY_n01_dpr450
        57.6%
        51%
        8.0
        HJMY5DRXY_n01_dpr451
        57.0%
        51%
        5.7
        HJMY5DRXY_n01_dpr452
        42.1%
        50%
        2.7
        HJMY5DRXY_n01_dpr453
        61.5%
        51%
        8.9
        HJMY5DRXY_n01_dpr454
        43.8%
        51%
        3.5
        HJMY5DRXY_n01_dpr455
        48.2%
        51%
        4.2
        HJMY5DRXY_n01_dpr456
        50.6%
        51%
        5.2
        HJMY5DRXY_n01_dpr457
        61.2%
        51%
        8.9
        HJMY5DRXY_n01_dpr458
        41.0%
        51%
        2.8
        HJMY5DRXY_n01_dpr459
        38.5%
        51%
        2.7
        HJMY5DRXY_n01_dpr460
        24.6%
        50%
        0.3
        HJMY5DRXY_n01_dpr461
        17.2%
        53%
        0.0
        HJMY5DRXY_n01_dpr462
        47.9%
        51%
        7.6
        HJMY5DRXY_n01_dpr463
        29.9%
        51%
        3.1
        HJMY5DRXY_n01_dpr464
        26.0%
        51%
        2.2
        HJMY5DRXY_n01_dpr465
        27.9%
        51%
        1.8
        HJMY5DRXY_n01_dpr466
        37.3%
        51%
        2.1
        HJMY5DRXY_n01_dpr467
        26.1%
        51%
        1.6
        HJMY5DRXY_n01_dpr468
        30.3%
        51%
        2.3
        HJMY5DRXY_n01_dpr469
        23.0%
        51%
        1.3
        HJMY5DRXY_n01_dpr470
        32.3%
        51%
        1.9
        HJMY5DRXY_n01_dpr471
        37.8%
        51%
        1.8
        HJMY5DRXY_n01_dpr472
        42.7%
        51%
        5.4
        HJMY5DRXY_n01_dpr473
        33.9%
        51%
        3.8
        HJMY5DRXY_n01_dpr474
        47.8%
        51%
        3.4
        HJMY5DRXY_n01_dpr475
        39.5%
        51%
        2.6
        HJMY5DRXY_n01_dpr476
        54.3%
        51%
        4.6
        HJMY5DRXY_n01_dpr477
        58.6%
        50%
        2.3
        HJMY5DRXY_n01_dpr478
        67.2%
        50%
        3.5
        HJMY5DRXY_n01_undetermined
        56.8%
        47%
        63.1

        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 78/78 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        63060835
        3.0
        dpr402
        33007798
        1.6
        dpr403
        35520872
        1.7
        dpr404
        53860178
        2.6
        dpr405
        30522424
        1.5
        dpr406
        32289330
        1.5
        dpr407
        60888427
        2.9
        dpr408
        33240913
        1.6
        dpr409
        29544588
        1.4
        dpr410
        713646577
        34.2
        dpr411
        36437189
        1.7
        dpr412
        28454948
        1.4
        dpr413
        32402379
        1.6
        dpr414
        25946119
        1.2
        dpr415
        36737796
        1.8
        dpr416
        34679988
        1.7
        dpr417
        37067650
        1.8
        dpr418
        37919678
        1.8
        dpr419
        40578569
        1.9
        dpr420
        41005880
        2.0
        dpr421
        34790766
        1.7
        dpr422
        35212141
        1.7
        dpr423
        40914179
        2.0
        dpr424
        36549565
        1.8
        dpr425
        36821041
        1.8
        dpr426
        25960058
        1.2
        dpr427
        30405104
        1.5
        dpr428
        28255145
        1.4
        dpr429
        28649664
        1.4
        dpr430
        26559605
        1.3
        dpr431
        29940009
        1.4
        dpr432
        28508154
        1.4
        dpr433
        24172987
        1.2
        dpr434
        28133105
        1.3
        dpr435
        31998964
        1.5
        dpr436
        28628428
        1.4
        dpr437
        29705966
        1.4
        dpr438
        1547908
        0.1
        dpr439
        357106
        0.0
        dpr440
        613826
        0.0
        dpr441
        485933
        0.0
        dpr442
        4581499
        0.2
        dpr443
        3293801
        0.2
        dpr444
        3532759
        0.2
        dpr445
        682.0
        0.0
        dpr446
        5568
        0.0
        dpr447
        2361213
        0.1
        dpr448
        1521278
        0.1
        dpr449
        1840662
        0.1
        dpr450
        7962679
        0.4
        dpr451
        5743778
        0.3
        dpr452
        2704611
        0.1
        dpr453
        8934431
        0.4
        dpr454
        3539499
        0.2
        dpr455
        4163990
        0.2
        dpr456
        5247850
        0.3
        dpr457
        8913644
        0.4
        dpr458
        2813262
        0.1
        dpr459
        2731715
        0.1
        dpr460
        307438
        0.0
        dpr461
        10850
        0.0
        dpr462
        7613177
        0.4
        dpr463
        3066802
        0.1
        dpr464
        2153872
        0.1
        dpr465
        1836867
        0.1
        dpr466
        2064085
        0.1
        dpr467
        1648127
        0.1
        dpr468
        2322342
        0.1
        dpr469
        1295969
        0.1
        dpr470
        1932547
        0.1
        dpr471
        1826432
        0.1
        dpr472
        5376368
        0.3
        dpr473
        3841252
        0.2
        dpr474
        3427098
        0.2
        dpr475
        2604852
        0.1
        dpr476
        4628095
        0.2
        dpr477
        2306546
        0.1
        dpr478
        3501547
        0.2

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        2.0
        2553348096
        2086678979
        3.0
        0.7

        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
        14552131.0
        23.1
        GGGGGGGGAGGCTATA
        2380953.0
        3.8
        CGGCTATGGGGGGGGG
        1315110.0
        2.1
        GGGGGGGGGCCTCTAT
        1130980.0
        1.8
        GGGGGGGGAGGATAGG
        912111.0
        1.4
        GGCTATGAAGGCTATA
        635491.0
        1.0
        CGCTATGAAGGCTATA
        502046.0
        0.8
        CGGCTATGACCAAAAA
        385435.0
        0.6
        CGGCTATGAGCTATAG
        326810.0
        0.5
        GGGGGGGGAGTACATC
        305396.0
        0.5
        CGGCTATGAGGCCTAT
        290205.0
        0.5
        CAAATATTAGGCTATA
        273595.0
        0.4
        CGGCTATGAGGCTTAT
        273277.0
        0.4
        CGGCTATGAGGGCTAT
        272155.0
        0.4
        CCGGCTATAGGCTATA
        256541.0
        0.4
        GGGGGGGGGAGTTAAG
        248790.0
        0.4
        CGGGCTATAGGCTATA
        231427.0
        0.4
        CGGCTATGTAGGCTAT
        226955.0
        0.4
        CGGCTATGGGCTATAG
        198668.0
        0.3
        GGGGGGGGAGTTCTCG
        190295.0
        0.3

        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..