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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 2022-07-14, 15:57 based on data in: /scratch/gencore/logs/html/HM3L5DSX3/merged


        General Statistics

        Showing 98/98 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HM3L5DSX3_n01_CG_1
        21.6%
        41%
        113.7
        HM3L5DSX3_n01_CG_10
        25.9%
        40%
        228.2
        HM3L5DSX3_n01_CG_11
        23.3%
        41%
        173.3
        HM3L5DSX3_n01_CG_12
        26.4%
        40%
        208.6
        HM3L5DSX3_n01_CG_13
        21.2%
        42%
        126.6
        HM3L5DSX3_n01_CG_14
        21.6%
        40%
        137.9
        HM3L5DSX3_n01_CG_15
        26.5%
        43%
        339.4
        HM3L5DSX3_n01_CG_16
        22.6%
        41%
        127.8
        HM3L5DSX3_n01_CG_17
        26.8%
        41%
        276.1
        HM3L5DSX3_n01_CG_18
        23.2%
        40%
        139.7
        HM3L5DSX3_n01_CG_19
        22.6%
        40%
        170.3
        HM3L5DSX3_n01_CG_2
        31.5%
        40%
        440.5
        HM3L5DSX3_n01_CG_20
        25.5%
        41%
        233.1
        HM3L5DSX3_n01_CG_21
        23.7%
        40%
        184.4
        HM3L5DSX3_n01_CG_22
        23.5%
        40%
        161.0
        HM3L5DSX3_n01_CG_23
        28.6%
        40%
        324.5
        HM3L5DSX3_n01_CG_24
        24.4%
        40%
        214.6
        HM3L5DSX3_n01_CG_25
        22.5%
        40%
        158.4
        HM3L5DSX3_n01_CG_26
        20.5%
        40%
        124.7
        HM3L5DSX3_n01_CG_27
        18.4%
        40%
        80.4
        HM3L5DSX3_n01_CG_28
        22.0%
        41%
        150.7
        HM3L5DSX3_n01_CG_29
        21.4%
        41%
        135.5
        HM3L5DSX3_n01_CG_3
        34.6%
        40%
        616.0
        HM3L5DSX3_n01_CG_30
        21.4%
        40%
        139.6
        HM3L5DSX3_n01_CG_31
        24.1%
        41%
        247.6
        HM3L5DSX3_n01_CG_32
        23.5%
        40%
        219.3
        HM3L5DSX3_n01_CG_33
        16.3%
        41%
        45.7
        HM3L5DSX3_n01_CG_34
        19.5%
        42%
        78.7
        HM3L5DSX3_n01_CG_35
        25.5%
        41%
        222.8
        HM3L5DSX3_n01_CG_36
        21.2%
        42%
        52.3
        HM3L5DSX3_n01_CG_37
        23.6%
        42%
        142.5
        HM3L5DSX3_n01_CG_38
        28.5%
        42%
        266.0
        HM3L5DSX3_n01_CG_39
        26.2%
        40%
        207.7
        HM3L5DSX3_n01_CG_4
        31.0%
        41%
        409.1
        HM3L5DSX3_n01_CG_40
        25.2%
        41%
        163.0
        HM3L5DSX3_n01_CG_41
        24.5%
        40%
        170.0
        HM3L5DSX3_n01_CG_42
        24.7%
        40%
        135.1
        HM3L5DSX3_n01_CG_43
        29.9%
        39%
        322.6
        HM3L5DSX3_n01_CG_44
        24.8%
        40%
        175.1
        HM3L5DSX3_n01_CG_45
        26.2%
        40%
        212.7
        HM3L5DSX3_n01_CG_46
        23.4%
        39%
        160.2
        HM3L5DSX3_n01_CG_47
        25.8%
        40%
        254.8
        HM3L5DSX3_n01_CG_48
        31.0%
        41%
        365.4
        HM3L5DSX3_n01_CG_5
        37.9%
        40%
        832.6
        HM3L5DSX3_n01_CG_6
        28.2%
        40%
        262.0
        HM3L5DSX3_n01_CG_7
        23.2%
        39%
        165.0
        HM3L5DSX3_n01_CG_8
        24.4%
        40%
        179.8
        HM3L5DSX3_n01_CG_9
        22.7%
        41%
        143.1
        HM3L5DSX3_n01_undetermined
        37.5%
        40%
        597.7
        HM3L5DSX3_n02_CG_1
        20.9%
        41%
        113.7
        HM3L5DSX3_n02_CG_10
        24.7%
        40%
        228.2
        HM3L5DSX3_n02_CG_11
        22.7%
        41%
        173.3
        HM3L5DSX3_n02_CG_12
        24.5%
        40%
        208.6
        HM3L5DSX3_n02_CG_13
        20.4%
        41%
        126.6
        HM3L5DSX3_n02_CG_14
        20.7%
        40%
        137.9
        HM3L5DSX3_n02_CG_15
        25.5%
        43%
        339.4
        HM3L5DSX3_n02_CG_16
        21.2%
        41%
        127.8
        HM3L5DSX3_n02_CG_17
        26.2%
        41%
        276.1
        HM3L5DSX3_n02_CG_18
        22.0%
        40%
        139.7
        HM3L5DSX3_n02_CG_19
        21.5%
        40%
        170.3
        HM3L5DSX3_n02_CG_2
        30.0%
        40%
        440.5
        HM3L5DSX3_n02_CG_20
        24.4%
        41%
        233.1
        HM3L5DSX3_n02_CG_21
        22.9%
        40%
        184.4
        HM3L5DSX3_n02_CG_22
        22.2%
        40%
        161.0
        HM3L5DSX3_n02_CG_23
        26.8%
        40%
        324.5
        HM3L5DSX3_n02_CG_24
        22.7%
        39%
        214.6
        HM3L5DSX3_n02_CG_25
        21.6%
        40%
        158.4
        HM3L5DSX3_n02_CG_26
        19.6%
        40%
        124.7
        HM3L5DSX3_n02_CG_27
        17.7%
        40%
        80.4
        HM3L5DSX3_n02_CG_28
        21.3%
        40%
        150.7
        HM3L5DSX3_n02_CG_29
        20.7%
        41%
        135.5
        HM3L5DSX3_n02_CG_3
        32.3%
        40%
        616.0
        HM3L5DSX3_n02_CG_30
        20.0%
        40%
        139.6
        HM3L5DSX3_n02_CG_31
        23.0%
        41%
        247.6
        HM3L5DSX3_n02_CG_32
        22.4%
        40%
        219.3
        HM3L5DSX3_n02_CG_33
        15.1%
        41%
        45.7
        HM3L5DSX3_n02_CG_34
        18.7%
        42%
        78.7
        HM3L5DSX3_n02_CG_35
        24.7%
        41%
        222.8
        HM3L5DSX3_n02_CG_36
        19.8%
        42%
        52.3
        HM3L5DSX3_n02_CG_37
        22.6%
        42%
        142.5
        HM3L5DSX3_n02_CG_38
        27.6%
        42%
        266.0
        HM3L5DSX3_n02_CG_39
        25.1%
        40%
        207.7
        HM3L5DSX3_n02_CG_4
        29.9%
        41%
        409.1
        HM3L5DSX3_n02_CG_40
        24.3%
        41%
        163.0
        HM3L5DSX3_n02_CG_41
        23.4%
        40%
        170.0
        HM3L5DSX3_n02_CG_42
        23.6%
        40%
        135.1
        HM3L5DSX3_n02_CG_43
        28.4%
        40%
        322.6
        HM3L5DSX3_n02_CG_44
        23.5%
        40%
        175.1
        HM3L5DSX3_n02_CG_45
        24.5%
        40%
        212.7
        HM3L5DSX3_n02_CG_46
        22.2%
        39%
        160.2
        HM3L5DSX3_n02_CG_47
        24.7%
        40%
        254.8
        HM3L5DSX3_n02_CG_48
        29.2%
        40%
        365.4
        HM3L5DSX3_n02_CG_5
        36.3%
        40%
        832.6
        HM3L5DSX3_n02_CG_6
        26.8%
        40%
        262.0
        HM3L5DSX3_n02_CG_7
        21.8%
        39%
        165.0
        HM3L5DSX3_n02_CG_8
        23.3%
        40%
        179.8
        HM3L5DSX3_n02_CG_9
        21.9%
        41%
        143.1
        HM3L5DSX3_n02_undetermined
        34.3%
        40%
        597.7

        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
        597713245
        5.4
        CG_1
        113661020
        1.0
        CG_2
        440547341
        4.0
        CG_3
        615995451
        5.6
        CG_4
        409065611
        3.7
        CG_5
        832632249
        7.5
        CG_6
        262026244
        2.4
        CG_7
        164973565
        1.5
        CG_8
        179837331
        1.6
        CG_9
        143130103
        1.3
        CG_10
        228204617
        2.1
        CG_11
        173253299
        1.6
        CG_12
        208577001
        1.9
        CG_13
        126612043
        1.1
        CG_14
        137871659
        1.2
        CG_15
        339445175
        3.1
        CG_16
        127821697
        1.2
        CG_17
        276127612
        2.5
        CG_18
        139744327
        1.3
        CG_19
        170250414
        1.5
        CG_20
        233097087
        2.1
        CG_21
        184408023
        1.7
        CG_22
        160995124
        1.5
        CG_23
        324507995
        2.9
        CG_24
        214619549
        1.9
        CG_25
        158427607
        1.4
        CG_26
        124717991
        1.1
        CG_27
        80404153
        0.7
        CG_28
        150707819
        1.4
        CG_29
        135469026
        1.2
        CG_30
        139583535
        1.3
        CG_31
        247599985
        2.2
        CG_32
        219321120
        2.0
        CG_33
        45661480
        0.4
        CG_34
        78699248
        0.7
        CG_35
        222808384
        2.0
        CG_36
        52298391
        0.5
        CG_37
        142454758
        1.3
        CG_38
        265987865
        2.4
        CG_39
        207664217
        1.9
        CG_40
        162987284
        1.5
        CG_41
        169978278
        1.5
        CG_42
        135092210
        1.2
        CG_43
        322554597
        2.9
        CG_44
        175114333
        1.6
        CG_45
        212732575
        1.9
        CG_46
        160213823
        1.5
        CG_47
        254756421
        2.3
        CG_48
        365404772
        3.3

        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
        11035757654
        5.4
        1.1

        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
        123221480.0
        20.6
        GTACCACAGGGTGTGG
        6153003.0
        1.0
        GGGGGGGGCAATCGAC
        3691244.0
        0.6
        GTACCACAGGGGGGGG
        2573579.0
        0.4
        GGGGGGGGCGATCTCG
        2276419.0
        0.4
        GGGGGGGGTAGTTGCG
        1754279.0
        0.3
        GGGGGGGGAAGCACTG
        1753423.0
        0.3
        GGGGGGGGTCAAGGAC
        1522840.0
        0.2
        TCGTCTGAGGGGGGGG
        1462330.0
        0.2
        GGGGGGGGTGATCTCG
        1306556.0
        0.2
        GTACCACAGGGGGTGG
        1280122.0
        0.2
        GGGGGGGGTGGTAGCT
        1171992.0
        0.2
        AAGACCGTGGGGGGGG
        1108969.0
        0.2
        GGGGGGGGGCAATGGA
        1102321.0
        0.2
        GTCCTAAGGGGGGGGG
        1090631.0
        0.2
        AAGACGTACAATCGAC
        1055458.0
        0.2
        TACCTGCAGGGGGGGG
        964424.0
        0.2
        GACGAACTGGGGGGGG
        936942.0
        0.2
        GGGGGGGGGTTCAACC
        935672.0
        0.2
        GGGGGGGGGGGGGGGG
        930262.0
        0.2

        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..