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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 2023-10-10, 02:37 based on data in: /scratch/gencore/logs/html/HL557DRX3/merged


        General Statistics

        Showing 110/110 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HL557DRX3_n01_Cisp01
        23.1%
        39%
        8.6
        HL557DRX3_n01_Cisp02
        29.2%
        41%
        8.8
        HL557DRX3_n01_Cisp03
        40.3%
        41%
        32.2
        HL557DRX3_n01_Cisp04
        30.5%
        41%
        16.1
        HL557DRX3_n01_Cisp05
        25.2%
        39%
        14.2
        HL557DRX3_n01_Cisp06
        26.2%
        39%
        10.5
        HL557DRX3_n01_Cisp07
        31.4%
        43%
        5.1
        HL557DRX3_n01_Cisp08
        24.8%
        39%
        15.3
        HL557DRX3_n01_Cisp09
        23.1%
        38%
        10.1
        HL557DRX3_n01_Cisp10
        23.1%
        40%
        9.6
        HL557DRX3_n01_Cisp11
        30.1%
        40%
        13.5
        HL557DRX3_n01_Cisp12
        22.3%
        38%
        11.2
        HL557DRX3_n01_Cisp13
        58.6%
        56%
        4.3
        HL557DRX3_n01_Cisp14
        37.9%
        41%
        66.1
        HL557DRX3_n01_Cisp15
        25.5%
        39%
        9.1
        HL557DRX3_n01_Cisp16
        24.3%
        39%
        12.1
        HL557DRX3_n01_Cisp17
        39.9%
        45%
        13.6
        HL557DRX3_n01_Cisp18
        21.7%
        38%
        10.4
        HL557DRX3_n01_Ctrl01
        19.0%
        37%
        9.1
        HL557DRX3_n01_Ctrl02
        21.3%
        38%
        10.9
        HL557DRX3_n01_Ctrl03
        37.0%
        46%
        8.8
        HL557DRX3_n01_Ctrl04
        52.3%
        55%
        1.0
        HL557DRX3_n01_Ctrl05
        24.2%
        40%
        8.7
        HL557DRX3_n01_Ctrl06
        22.7%
        38%
        10.2
        HL557DRX3_n01_Ctrl07
        22.6%
        41%
        4.8
        HL557DRX3_n01_Ctrl08
        23.8%
        42%
        6.1
        HL557DRX3_n01_Ctrl09
        27.0%
        42%
        6.7
        HL557DRX3_n01_Ctrl10
        23.7%
        41%
        6.4
        HL557DRX3_n01_Ctrl11
        22.9%
        40%
        5.9
        HL557DRX3_n01_Ctrl12
        24.0%
        40%
        7.3
        HL557DRX3_n01_Ctrl13
        27.3%
        42%
        3.9
        HL557DRX3_n01_Ctrl14
        27.1%
        43%
        5.0
        HL557DRX3_n01_Ctrl15
        22.4%
        41%
        5.1
        HL557DRX3_n01_Ctrl16
        22.1%
        39%
        6.7
        HL557DRX3_n01_Ctrl17
        28.0%
        43%
        5.9
        HL557DRX3_n01_Ctrl18
        21.9%
        41%
        7.0
        HL557DRX3_n01_EMS01
        21.1%
        39%
        9.5
        HL557DRX3_n01_EMS02
        20.5%
        40%
        7.6
        HL557DRX3_n01_EMS03
        49.6%
        47%
        41.5
        HL557DRX3_n01_EMS04
        64.7%
        51%
        95.3
        HL557DRX3_n01_EMS05
        26.2%
        38%
        18.4
        HL557DRX3_n01_EMS06
        24.4%
        40%
        8.7
        HL557DRX3_n01_EMS07
        25.9%
        38%
        16.8
        HL557DRX3_n01_EMS08
        24.4%
        40%
        10.0
        HL557DRX3_n01_EMS09
        26.5%
        38%
        19.2
        HL557DRX3_n01_EMS10
        24.5%
        40%
        6.4
        HL557DRX3_n01_EMS11
        29.0%
        40%
        9.1
        HL557DRX3_n01_EMS12
        23.4%
        41%
        4.5
        HL557DRX3_n01_EMS13
        22.5%
        40%
        8.9
        HL557DRX3_n01_EMS14
        24.0%
        39%
        15.1
        HL557DRX3_n01_EMS15
        21.3%
        38%
        9.6
        HL557DRX3_n01_EMS16
        24.2%
        42%
        3.9
        HL557DRX3_n01_EMS17
        20.0%
        39%
        9.0
        HL557DRX3_n01_EMS18
        21.3%
        39%
        9.5
        HL557DRX3_n01_undetermined
        77.7%
        41%
        1067.9
        HL557DRX3_n02_Cisp01
        21.4%
        40%
        8.6
        HL557DRX3_n02_Cisp02
        27.1%
        43%
        8.8
        HL557DRX3_n02_Cisp03
        38.8%
        42%
        32.2
        HL557DRX3_n02_Cisp04
        29.3%
        42%
        16.1
        HL557DRX3_n02_Cisp05
        24.5%
        40%
        14.2
        HL557DRX3_n02_Cisp06
        24.8%
        40%
        10.5
        HL557DRX3_n02_Cisp07
        28.4%
        45%
        5.1
        HL557DRX3_n02_Cisp08
        24.0%
        39%
        15.3
        HL557DRX3_n02_Cisp09
        20.9%
        39%
        10.1
        HL557DRX3_n02_Cisp10
        21.8%
        40%
        9.6
        HL557DRX3_n02_Cisp11
        28.9%
        41%
        13.5
        HL557DRX3_n02_Cisp12
        21.4%
        39%
        11.2
        HL557DRX3_n02_Cisp13
        53.2%
        64%
        4.3
        HL557DRX3_n02_Cisp14
        36.0%
        42%
        66.1
        HL557DRX3_n02_Cisp15
        23.6%
        40%
        9.1
        HL557DRX3_n02_Cisp16
        23.4%
        40%
        12.1
        HL557DRX3_n02_Cisp17
        37.3%
        48%
        13.6
        HL557DRX3_n02_Cisp18
        20.5%
        38%
        10.4
        HL557DRX3_n02_Ctrl01
        17.8%
        38%
        9.1
        HL557DRX3_n02_Ctrl02
        20.2%
        38%
        10.9
        HL557DRX3_n02_Ctrl03
        33.9%
        50%
        8.8
        HL557DRX3_n02_Ctrl04
        48.0%
        62%
        1.0
        HL557DRX3_n02_Ctrl05
        22.9%
        40%
        8.7
        HL557DRX3_n02_Ctrl06
        21.2%
        38%
        10.2
        HL557DRX3_n02_Ctrl07
        21.2%
        42%
        4.8
        HL557DRX3_n02_Ctrl08
        22.6%
        42%
        6.1
        HL557DRX3_n02_Ctrl09
        25.9%
        43%
        6.7
        HL557DRX3_n02_Ctrl10
        22.5%
        42%
        6.4
        HL557DRX3_n02_Ctrl11
        20.9%
        41%
        5.9
        HL557DRX3_n02_Ctrl12
        21.9%
        40%
        7.3
        HL557DRX3_n02_Ctrl13
        25.4%
        44%
        3.9
        HL557DRX3_n02_Ctrl14
        25.6%
        45%
        5.0
        HL557DRX3_n02_Ctrl15
        20.9%
        41%
        5.1
        HL557DRX3_n02_Ctrl16
        21.0%
        40%
        6.7
        HL557DRX3_n02_Ctrl17
        26.3%
        45%
        5.9
        HL557DRX3_n02_Ctrl18
        19.4%
        41%
        7.0
        HL557DRX3_n02_EMS01
        20.3%
        39%
        9.5
        HL557DRX3_n02_EMS02
        19.4%
        41%
        7.6
        HL557DRX3_n02_EMS03
        47.0%
        50%
        41.5
        HL557DRX3_n02_EMS04
        61.9%
        56%
        95.3
        HL557DRX3_n02_EMS05
        25.1%
        38%
        18.4
        HL557DRX3_n02_EMS06
        23.1%
        41%
        8.7
        HL557DRX3_n02_EMS07
        25.1%
        39%
        16.8
        HL557DRX3_n02_EMS08
        22.4%
        41%
        10.0
        HL557DRX3_n02_EMS09
        25.7%
        38%
        19.2
        HL557DRX3_n02_EMS10
        22.9%
        41%
        6.4
        HL557DRX3_n02_EMS11
        27.2%
        42%
        9.1
        HL557DRX3_n02_EMS12
        22.1%
        43%
        4.5
        HL557DRX3_n02_EMS13
        21.5%
        40%
        8.9
        HL557DRX3_n02_EMS14
        23.3%
        39%
        15.1
        HL557DRX3_n02_EMS15
        20.3%
        38%
        9.6
        HL557DRX3_n02_EMS16
        22.3%
        44%
        3.9
        HL557DRX3_n02_EMS17
        19.1%
        39%
        9.0
        HL557DRX3_n02_EMS18
        20.1%
        40%
        9.5
        HL557DRX3_n02_undetermined
        73.5%
        42%
        1067.9

        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 55/55 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        1067943592
        60.6
        EMS01
        9461256
        0.5
        EMS02
        7634548
        0.4
        EMS03
        41541874
        2.4
        EMS04
        95251767
        5.4
        EMS05
        18434810
        1.0
        EMS06
        8743091
        0.5
        EMS07
        16838719
        1.0
        EMS08
        10030291
        0.6
        EMS09
        19230892
        1.1
        EMS10
        6354039
        0.4
        EMS11
        9076042
        0.5
        EMS12
        4478326
        0.3
        EMS13
        8885255
        0.5
        EMS14
        15098854
        0.9
        EMS15
        9557667
        0.5
        EMS16
        3869298
        0.2
        EMS17
        9040206
        0.5
        EMS18
        9481526
        0.5
        Cisp01
        8623236
        0.5
        Cisp02
        8784279
        0.5
        Cisp03
        32168999
        1.8
        Cisp04
        16125665
        0.9
        Cisp05
        14159772
        0.8
        Cisp06
        10473084
        0.6
        Cisp07
        5120101
        0.3
        Cisp08
        15294677
        0.9
        Cisp09
        10067404
        0.6
        Cisp10
        9556726
        0.5
        Cisp11
        13536995
        0.8
        Cisp12
        11188524
        0.6
        Cisp13
        4262441
        0.2
        Cisp14
        66110657
        3.8
        Cisp15
        9112245
        0.5
        Cisp16
        12108232
        0.7
        Cisp17
        13640705
        0.8
        Cisp18
        10428164
        0.6
        Ctrl01
        9075741
        0.5
        Ctrl02
        10907738
        0.6
        Ctrl03
        8790100
        0.5
        Ctrl04
        999392
        0.1
        Ctrl05
        8652349
        0.5
        Ctrl06
        10208178
        0.6
        Ctrl07
        4818044
        0.3
        Ctrl08
        6128972
        0.3
        Ctrl09
        6675800
        0.4
        Ctrl10
        6400843
        0.4
        Ctrl11
        5921815
        0.3
        Ctrl12
        7250116
        0.4
        Ctrl13
        3870916
        0.2
        Ctrl14
        4997551
        0.3
        Ctrl15
        5135314
        0.3
        Ctrl16
        6722459
        0.4
        Ctrl17
        5902244
        0.3
        Ctrl18
        7040364
        0.4

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

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGGGGAGATCTCG
        71419157.0
        6.7
        CGAATACGTCAGACGA
        21291711.0
        2.0
        GTCCTTGAGTCGGTAA
        21244277.0
        2.0
        GTCCTTGAGGTGTCTT
        16015436.0
        1.5
        GTAACCGAGTCGGTAA
        15495782.0
        1.4
        CGAATACGGGTGTCTT
        15346190.0
        1.4
        GTAACCGATCAGACGA
        13710430.0
        1.3
        CAGTGCTTGTCGGTAA
        11118863.0
        1.0
        CAGTGCTTTCAGACGA
        9669071.0
        0.9
        GGGGGGGGTCAGACGA
        6467547.0
        0.6
        GGGGGGGGGTCGGTAA
        6087840.0
        0.6
        CGAATACGACCTGGAA
        6065432.0
        0.6
        GTCCTTGAACCTGGAA
        5759232.0
        0.5
        TCCATTGCTCAGACGA
        5126118.0
        0.5
        TCCATTGCGTCGGTAA
        4873237.0
        0.5
        TAGGAGCTGTCGGTAA
        4803651.0
        0.5
        GTCCTTGAAGACCGTA
        4548231.0
        0.4
        CGAATACGAGATCTCG
        4227246.0
        0.4
        TAGGAGCTTCAGACGA
        4095219.0
        0.4
        CGAATACGAGACCGTA
        4021138.0
        0.4

        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
        1761211895
        60.6
        5.6

        FastQC

        FastQC is a quality control tool for high throughput sequence data, written by Simon Andrews at the Babraham Institute in Cambridge.

        Sequence Counts

        Sequence counts for each sample. Duplicate read counts are an estimate only.

        This plot show the total number of reads, broken down into unique and duplicate if possible (only more recent versions of FastQC give duplicate info).

        You can read more about duplicate calculation in the FastQC documentation. A small part has been copied here for convenience:

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Quality Histograms

        The mean quality value across each base position in the read.

        To enable multiple samples to be plotted on the same graph, only the mean quality scores are plotted (unlike the box plots seen in FastQC reports).

        Taken from the FastQC help:

        The y-axis on the graph shows the quality scores. The higher the score, the better the base call. The background of the graph divides the y axis into very good quality calls (green), calls of reasonable quality (orange), and calls of poor quality (red). The quality of calls on most platforms will degrade as the run progresses, so it is common to see base calls falling into the orange area towards the end of a read.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Sequence Quality Scores

        The number of reads with average quality scores. Shows if a subset of reads has poor quality.

        From the FastQC help:

        The per sequence quality score report allows you to see if a subset of your sequences have universally low quality values. It is often the case that a subset of sequences will have universally poor quality, however these should represent only a small percentage of the total sequences.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Base Sequence Content

        The proportion of each base position for which each of the four normal DNA bases has been called.

        To enable multiple samples to be shown in a single plot, the base composition data is shown as a heatmap. The colours represent the balance between the four bases: an even distribution should give an even muddy brown colour. Hover over the plot to see the percentage of the four bases under the cursor.

        To see the data as a line plot, as in the original FastQC graph, click on a sample track.

        From the FastQC help:

        Per Base Sequence Content plots out the proportion of each base position in a file for which each of the four normal DNA bases has been called.

        In a random library you would expect that there would be little to no difference between the different bases of a sequence run, so the lines in this plot should run parallel with each other. The relative amount of each base should reflect the overall amount of these bases in your genome, but in any case they should not be hugely imbalanced from each other.

        It's worth noting that some types of library will always produce biased sequence composition, normally at the start of the read. Libraries produced by priming using random hexamers (including nearly all RNA-Seq libraries) and those which were fragmented using transposases inherit an intrinsic bias in the positions at which reads start. This bias does not concern an absolute sequence, but instead provides enrichement of a number of different K-mers at the 5' end of the reads. Whilst this is a true technical bias, it isn't something which can be corrected by trimming and in most cases doesn't seem to adversely affect the downstream analysis.

        Click a sample row to see a line plot for that dataset.
        Rollover for sample name
        Position: -
        %T: -
        %C: -
        %A: -
        %G: -

        Per Sequence GC Content

        The average GC content of reads. Normal random library typically have a roughly normal distribution of GC content.

        From the FastQC help:

        This module measures the GC content across the whole length of each sequence in a file and compares it to a modelled normal distribution of GC content.

        In a normal random library you would expect to see a roughly normal distribution of GC content where the central peak corresponds to the overall GC content of the underlying genome. Since we don't know the the GC content of the genome the modal GC content is calculated from the observed data and used to build a reference distribution.

        An unusually shaped distribution could indicate a contaminated library or some other kinds of biased subset. A normal distribution which is shifted indicates some systematic bias which is independent of base position. If there is a systematic bias which creates a shifted normal distribution then this won't be flagged as an error by the module since it doesn't know what your genome's GC content should be.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Per Base N Content

        The percentage of base calls at each position for which an N was called.

        From the FastQC help:

        If a sequencer is unable to make a base call with sufficient confidence then it will normally substitute an N rather than a conventional base call. This graph shows the percentage of base calls at each position for which an N was called.

        It's not unusual to see a very low proportion of Ns appearing in a sequence, especially nearer the end of a sequence. However, if this proportion rises above a few percent it suggests that the analysis pipeline was unable to interpret the data well enough to make valid base calls.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Length Distribution

        All samples have sequences of a single length (151bp).

        Sequence Duplication Levels

        The relative level of duplication found for every sequence.

        From the FastQC Help:

        In a diverse library most sequences will occur only once in the final set. A low level of duplication may indicate a very high level of coverage of the target sequence, but a high level of duplication is more likely to indicate some kind of enrichment bias (eg PCR over amplification). This graph shows the degree of duplication for every sequence in a library: the relative number of sequences with different degrees of duplication.

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        In a properly diverse library most sequences should fall into the far left of the plot in both the red and blue lines. A general level of enrichment, indicating broad oversequencing in the library will tend to flatten the lines, lowering the low end and generally raising other categories. More specific enrichments of subsets, or the presence of low complexity contaminants will tend to produce spikes towards the right of the plot.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Overrepresented sequences

        The total amount of overrepresented sequences found in each library.

        FastQC calculates and lists overrepresented sequences in FastQ files. It would not be possible to show this for all samples in a MultiQC report, so instead this plot shows the number of sequences categorized as over represented.

        Sometimes, a single sequence may account for a large number of reads in a dataset. To show this, the bars are split into two: the first shows the overrepresented reads that come from the single most common sequence. The second shows the total count from all remaining overrepresented sequences.

        From the FastQC Help:

        A normal high-throughput library will contain a diverse set of sequences, with no individual sequence making up a tiny fraction of the whole. Finding that a single sequence is very overrepresented in the set either means that it is highly biologically significant, or indicates that the library is contaminated, or not as diverse as you expected.

        FastQC lists all of the sequences which make up more than 0.1% of the total. To conserve memory only sequences which appear in the first 100,000 sequences are tracked to the end of the file. It is therefore possible that a sequence which is overrepresented but doesn't appear at the start of the file for some reason could be missed by this module.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Adapter Content

        The cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position.

        Note that only samples with ≥ 0.1% adapter contamination are shown.

        There may be several lines per sample, as one is shown for each adapter detected in the file.

        From the FastQC Help:

        The plot shows a cumulative percentage count of the proportion of your library which has seen each of the adapter sequences at each position. Once a sequence has been seen in a read it is counted as being present right through to the end of the read so the percentages you see will only increase as the read length goes on.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Status Checks

        Status for each FastQC section showing whether results seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        FastQC assigns a status for each section of the report. These give a quick evaluation of whether the results of the analysis seem entirely normal (green), slightly abnormal (orange) or very unusual (red).

        It is important to stress that although the analysis results appear to give a pass/fail result, these evaluations must be taken in the context of what you expect from your library. A 'normal' sample as far as FastQC is concerned is random and diverse. Some experiments may be expected to produce libraries which are biased in particular ways. You should treat the summary evaluations therefore as pointers to where you should concentrate your attention and understand why your library may not look random and diverse.

        Specific guidance on how to interpret the output of each module can be found in the relevant report section, or in the FastQC help.

        In this heatmap, we summarise all of these into a single heatmap for a quick overview. Note that not all FastQC sections have plots in MultiQC reports, but all status checks are shown in this heatmap.

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