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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 2022-03-23, 05:50 based on data in: /scratch/gencore/logs/html/HMYYNDRXY/1


        General Statistics

        Showing 97/97 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HMYYNDRXY_l01_n01_RS100J2
        28.7%
        38%
        9.8
        HMYYNDRXY_l01_n01_RS101J2
        28.4%
        38%
        9.2
        HMYYNDRXY_l01_n01_RS102J2
        28.1%
        39%
        9.4
        HMYYNDRXY_l01_n01_RS103J2
        27.4%
        38%
        9.5
        HMYYNDRXY_l01_n01_RS106J2
        23.2%
        37%
        9.7
        HMYYNDRXY_l01_n01_RS107J2
        28.7%
        39%
        9.2
        HMYYNDRXY_l01_n01_RS108J2
        26.5%
        40%
        8.4
        HMYYNDRXY_l01_n01_RS109J2
        28.8%
        40%
        8.9
        HMYYNDRXY_l01_n01_RS110J2
        41.0%
        39%
        8.7
        HMYYNDRXY_l01_n01_RS111J2
        25.5%
        39%
        7.7
        HMYYNDRXY_l01_n01_RS112J2
        23.2%
        38%
        8.9
        HMYYNDRXY_l01_n01_RS113J2
        24.8%
        39%
        10.2
        HMYYNDRXY_l01_n01_RS114J2
        27.1%
        39%
        12.6
        HMYYNDRXY_l01_n01_RS115J2
        28.5%
        38%
        17.0
        HMYYNDRXY_l01_n01_RS116J2
        29.6%
        38%
        19.1
        HMYYNDRXY_l01_n01_RS117J2
        23.0%
        38%
        10.1
        HMYYNDRXY_l01_n01_RS118J2
        24.3%
        38%
        9.0
        HMYYNDRXY_l01_n01_RS119J2
        23.0%
        37%
        9.3
        HMYYNDRXY_l01_n01_RS120J2
        23.0%
        38%
        9.7
        HMYYNDRXY_l01_n01_RS121J2
        20.1%
        38%
        7.7
        HMYYNDRXY_l01_n01_RS122J2
        24.7%
        38%
        10.3
        HMYYNDRXY_l01_n01_RS123J2
        22.5%
        37%
        10.1
        HMYYNDRXY_l01_n01_RS124J2
        23.9%
        37%
        9.5
        HMYYNDRXY_l01_n01_RS125J2
        23.4%
        37%
        10.2
        HMYYNDRXY_l01_n01_RS126J2
        22.9%
        37%
        7.7
        HMYYNDRXY_l01_n01_RS127J2
        19.2%
        37%
        5.0
        HMYYNDRXY_l01_n01_RS128J2
        25.6%
        38%
        10.6
        HMYYNDRXY_l01_n01_RS129J2
        22.2%
        37%
        7.5
        HMYYNDRXY_l01_n01_RS130J2
        28.3%
        38%
        11.3
        HMYYNDRXY_l01_n01_RS131J2
        23.8%
        38%
        10.6
        HMYYNDRXY_l01_n01_RS132J2
        23.8%
        37%
        10.4
        HMYYNDRXY_l01_n01_RS133J2
        26.3%
        38%
        10.1
        HMYYNDRXY_l01_n01_RS134J2
        32.8%
        39%
        9.6
        HMYYNDRXY_l01_n01_RS135J2
        23.4%
        37%
        10.4
        HMYYNDRXY_l01_n01_RS136J2
        22.5%
        36%
        10.7
        HMYYNDRXY_l01_n01_RS137J2
        31.4%
        38%
        10.1
        HMYYNDRXY_l01_n01_RS138J2
        31.6%
        37%
        10.3
        HMYYNDRXY_l01_n01_RS139J2
        27.2%
        39%
        8.2
        HMYYNDRXY_l01_n01_RS140J2
        18.5%
        37%
        4.9
        HMYYNDRXY_l01_n01_RS141J2
        24.2%
        37%
        8.9
        HMYYNDRXY_l01_n01_RS142J2
        25.1%
        37%
        11.2
        HMYYNDRXY_l01_n01_RS143J2
        22.5%
        37%
        8.7
        HMYYNDRXY_l01_n01_RS144J2
        23.7%
        37%
        11.5
        HMYYNDRXY_l01_n01_RS145J2
        23.4%
        38%
        10.7
        HMYYNDRXY_l01_n01_RS146J2
        31.3%
        37%
        23.7
        HMYYNDRXY_l01_n01_RS49J2
        25.6%
        38%
        10.0
        HMYYNDRXY_l01_n01_RS50J2
        29.4%
        40%
        9.6
        HMYYNDRXY_l01_n01_RS51J2
        25.4%
        38%
        9.6
        HMYYNDRXY_l01_n01_RS52J2
        25.2%
        39%
        8.4
        HMYYNDRXY_l01_n01_RS53J2
        22.1%
        37%
        9.0
        HMYYNDRXY_l01_n01_RS54J2
        22.2%
        37%
        9.5
        HMYYNDRXY_l01_n01_RS55J2
        22.9%
        37%
        8.9
        HMYYNDRXY_l01_n01_RS56J2
        21.5%
        37%
        11.0
        HMYYNDRXY_l01_n01_RS57J2
        16.8%
        37%
        4.7
        HMYYNDRXY_l01_n01_RS58J2
        22.9%
        37%
        9.6
        HMYYNDRXY_l01_n01_RS59J2
        17.3%
        37%
        4.0
        HMYYNDRXY_l01_n01_RS60J2
        22.3%
        37%
        10.7
        HMYYNDRXY_l01_n01_RS61J2
        24.0%
        39%
        8.9
        HMYYNDRXY_l01_n01_RS62J2
        26.8%
        38%
        9.2
        HMYYNDRXY_l01_n01_RS63J2
        20.3%
        38%
        9.1
        HMYYNDRXY_l01_n01_RS64J2
        22.2%
        37%
        9.1
        HMYYNDRXY_l01_n01_RS65J2
        21.1%
        37%
        10.0
        HMYYNDRXY_l01_n01_RS66J2
        18.2%
        37%
        4.1
        HMYYNDRXY_l01_n01_RS67J2
        22.8%
        38%
        10.0
        HMYYNDRXY_l01_n01_RS68J2
        22.9%
        37%
        9.3
        HMYYNDRXY_l01_n01_RS69J2
        19.2%
        38%
        4.3
        HMYYNDRXY_l01_n01_RS70J2
        23.7%
        37%
        10.4
        HMYYNDRXY_l01_n01_RS71J2
        31.3%
        38%
        9.5
        HMYYNDRXY_l01_n01_RS72J2
        24.6%
        37%
        8.1
        HMYYNDRXY_l01_n01_RS73J2
        24.5%
        37%
        8.7
        HMYYNDRXY_l01_n01_RS74J2
        25.2%
        38%
        9.4
        HMYYNDRXY_l01_n01_RS75J2
        32.5%
        36%
        8.2
        HMYYNDRXY_l01_n01_RS76J2
        22.5%
        37%
        9.4
        HMYYNDRXY_l01_n01_RS77J2
        21.3%
        37%
        7.4
        HMYYNDRXY_l01_n01_RS78J2
        24.3%
        38%
        8.3
        HMYYNDRXY_l01_n01_RS79J2
        22.1%
        38%
        8.2
        HMYYNDRXY_l01_n01_RS80J2
        24.8%
        37%
        8.9
        HMYYNDRXY_l01_n01_RS81J2
        24.0%
        39%
        8.9
        HMYYNDRXY_l01_n01_RS82J2
        29.8%
        38%
        9.0
        HMYYNDRXY_l01_n01_RS83J2
        25.6%
        38%
        8.9
        HMYYNDRXY_l01_n01_RS84J2
        32.4%
        39%
        7.5
        HMYYNDRXY_l01_n01_RS85J2
        16.2%
        37%
        3.6
        HMYYNDRXY_l01_n01_RS86J2
        24.7%
        38%
        7.7
        HMYYNDRXY_l01_n01_RS87J2
        22.2%
        37%
        8.8
        HMYYNDRXY_l01_n01_RS88J2
        28.6%
        38%
        6.2
        HMYYNDRXY_l01_n01_RS89J2
        22.2%
        38%
        9.5
        HMYYNDRXY_l01_n01_RS90J2
        26.0%
        38%
        9.1
        HMYYNDRXY_l01_n01_RS91J2
        21.0%
        38%
        7.7
        HMYYNDRXY_l01_n01_RS92J2
        24.1%
        38%
        7.3
        HMYYNDRXY_l01_n01_RS93J2
        23.9%
        38%
        10.1
        HMYYNDRXY_l01_n01_RS94J2
        24.8%
        38%
        9.3
        HMYYNDRXY_l01_n01_RS95J2
        24.0%
        38%
        8.2
        HMYYNDRXY_l01_n01_RS96J2
        25.7%
        37%
        8.9
        HMYYNDRXY_l01_n01_RS97J2
        23.4%
        38%
        9.5
        HMYYNDRXY_l01_n01_RS98J2
        25.6%
        38%
        8.5
        HMYYNDRXY_l01_n01_RS99J2
        23.0%
        38%
        8.7
        HMYYNDRXY_l01_n01_undetermined
        95.2%
        44%
        215.5

        Lane 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 97/97 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        215524636
        19.5
        RS49J2
        9960118
        0.9
        RS50J2
        9626869
        0.9
        RS51J2
        9580375
        0.9
        RS52J2
        8410463
        0.8
        RS53J2
        9048268
        0.8
        RS54J2
        9470572
        0.9
        RS55J2
        8916052
        0.8
        RS56J2
        10999375
        1.0
        RS57J2
        4713911
        0.4
        RS58J2
        9578830
        0.9
        RS59J2
        3987405
        0.4
        RS60J2
        10699399
        1.0
        RS61J2
        8909494
        0.8
        RS62J2
        9157729
        0.8
        RS63J2
        9095197
        0.8
        RS64J2
        9074235
        0.8
        RS65J2
        9990024
        0.9
        RS66J2
        4099136
        0.4
        RS67J2
        9954872
        0.9
        RS68J2
        9254741
        0.8
        RS69J2
        4300752
        0.4
        RS70J2
        10392234
        0.9
        RS71J2
        9465531
        0.9
        RS72J2
        8113887
        0.7
        RS73J2
        8741789
        0.8
        RS74J2
        9370326
        0.8
        RS75J2
        8205207
        0.7
        RS76J2
        9404383
        0.9
        RS77J2
        7391246
        0.7
        RS78J2
        8286358
        0.8
        RS79J2
        8241968
        0.7
        RS80J2
        8942353
        0.8
        RS81J2
        8869490
        0.8
        RS82J2
        8952680
        0.8
        RS83J2
        8852454
        0.8
        RS84J2
        7523286
        0.7
        RS85J2
        3580261
        0.3
        RS86J2
        7671352
        0.7
        RS87J2
        8764459
        0.8
        RS88J2
        6200283
        0.6
        RS89J2
        9493098
        0.9
        RS90J2
        9073853
        0.8
        RS91J2
        7671612
        0.7
        RS92J2
        7274824
        0.7
        RS93J2
        10131774
        0.9
        RS94J2
        9341195
        0.8
        RS95J2
        8193467
        0.7
        RS96J2
        8949895
        0.8
        RS97J2
        9507541
        0.9
        RS98J2
        8519079
        0.8
        RS99J2
        8709840
        0.8
        RS100J2
        9833154
        0.9
        RS101J2
        9207944
        0.8
        RS102J2
        9408927
        0.9
        RS103J2
        9513868
        0.9
        RS106J2
        9717575
        0.9
        RS107J2
        9233002
        0.8
        RS108J2
        8389837
        0.8
        RS109J2
        8876393
        0.8
        RS110J2
        8678774
        0.8
        RS111J2
        7736266
        0.7
        RS112J2
        8947087
        0.8
        RS113J2
        10228427
        0.9
        RS114J2
        12557319
        1.1
        RS115J2
        16968406
        1.5
        RS116J2
        19055001
        1.7
        RS117J2
        10117860
        0.9
        RS118J2
        9002535
        0.8
        RS119J2
        9287359
        0.8
        RS120J2
        9682556
        0.9
        RS121J2
        7735215
        0.7
        RS122J2
        10310840
        0.9
        RS123J2
        10101666
        0.9
        RS124J2
        9476121
        0.9
        RS125J2
        10232465
        0.9
        RS126J2
        7716511
        0.7
        RS127J2
        5041357
        0.5
        RS128J2
        10559648
        1.0
        RS129J2
        7492993
        0.7
        RS130J2
        11256378
        1.0
        RS131J2
        10584043
        1.0
        RS132J2
        10441478
        0.9
        RS133J2
        10097896
        0.9
        RS134J2
        9605983
        0.9
        RS135J2
        10366479
        0.9
        RS136J2
        10700855
        1.0
        RS137J2
        10074726
        0.9
        RS138J2
        10298495
        0.9
        RS139J2
        8194522
        0.7
        RS140J2
        4911175
        0.4
        RS141J2
        8857926
        0.8
        RS142J2
        11152962
        1.0
        RS143J2
        8701502
        0.8
        RS144J2
        11513550
        1.0
        RS145J2
        10698615
        1.0
        RS146J2
        23701465
        2.1

        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 (%)
        GGGGGG
        210374931.0
        97.6
        CGGGGG
        96725.0
        0.0
        AAAAAA
        86685.0
        0.0
        NGGGGG
        62972.0
        0.0
        GGGGGC
        55001.0
        0.0
        AAACAA
        45539.0
        0.0
        TTATGA
        44963.0
        0.0
        ACAAAA
        40490.0
        0.0
        TGGTAA
        39633.0
        0.0
        AAAAAC
        38920.0
        0.0
        TGATTA
        38524.0
        0.0
        CAAAAA
        38490.0
        0.0
        AAAATA
        36525.0
        0.0
        GGGGCG
        36399.0
        0.0
        GGGGTG
        34104.0
        0.0
        TTGTAA
        32155.0
        0.0
        AAATAA
        31301.0
        0.0
        AAAAAT
        30373.0
        0.0
        GNGGGG
        30051.0
        0.0
        ATTGTA
        29931.0
        0.0

        Lane Statistics

        Showing 1/1 rows and 4/4 columns.
        LaneTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        1.0
        1276674048
        1104455334
        19.5
        18.6

        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.

        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.

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        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.

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        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.

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        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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