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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 2023-03-25, 08:27 based on data in: /scratch/gencore/logs/html/000000000-DKV2Y/1


        General Statistics

        Showing 526 samples.

        loading..

        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 263/263 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        106810
        6.8
        A01_ATH
        3861
        0.2
        B01_ATH
        2896
        0.2
        C01_ATH
        4559
        0.3
        D01_ATH
        391.0
        0.0
        E01_ATH
        2292
        0.1
        F01_ATH
        4260
        0.3
        G01_ATH
        6183
        0.4
        H01_ATH
        3932
        0.3
        A02_ATH
        5409
        0.3
        B02_ATH
        4039
        0.3
        C02_ATH
        5948
        0.4
        D02_ATH
        5019
        0.3
        E02_ATH
        3640
        0.2
        F02_ATH
        1912
        0.1
        G02_ATH
        8108
        0.5
        H02_ATH
        7225
        0.5
        A03_ATH
        5857
        0.4
        B03_ATH
        4806
        0.3
        C03_ATH
        4150
        0.3
        D03_ATH
        3467
        0.2
        E03_ATH
        4136
        0.3
        F03_ATH
        2660
        0.2
        G03_ATH
        8774
        0.6
        H03_ATH
        6190
        0.4
        A04_ATH
        6.0
        0.0
        B04_ATH
        9.0
        0.0
        C04_ATH
        14.0
        0.0
        D04_ATH
        11.0
        0.0
        E04_ATH
        7.0
        0.0
        F04_ATH
        49.0
        0.0
        G04_ATH
        18.0
        0.0
        H04_ATH
        18.0
        0.0
        A05_ATH
        1050
        0.1
        B05_ATH
        1757
        0.1
        C05_ATH
        2404
        0.2
        D05_ATH
        2226
        0.1
        E05_ATH
        1977
        0.1
        F05_ATH
        2486
        0.2
        G05_ATH
        3177
        0.2
        H05_ATH
        2992
        0.2
        A06_ATH
        4781
        0.3
        B06_ATH
        2285
        0.1
        C06_ATH
        1786
        0.1
        D06_ATH
        3307
        0.2
        E06_ATH
        5907
        0.4
        F06_ATH
        2906
        0.2
        G06_ATH
        5948
        0.4
        H06_ATH
        3623
        0.2
        A07_ATH
        6665
        0.4
        B07_ATH
        5099
        0.3
        C07_ATH
        3007
        0.2
        D07_ATH
        5579
        0.4
        E07_ATH
        3245
        0.2
        F07_ATH
        9826
        0.6
        G07_ATH
        10846
        0.7
        H07_ATH
        7532
        0.5
        A08_ATH
        6403
        0.4
        B08_ATH
        4383
        0.3
        C08_ATH
        6147
        0.4
        D08_ATH
        10756
        0.7
        E08_ATH
        2708
        0.2
        F08_ATH
        8285
        0.5
        G08_ATH
        9891
        0.6
        H08_ATH
        9953
        0.6
        A09_ATH
        13.0
        0.0
        B09_ATH
        7.0
        0.0
        C09_ATH
        7.0
        0.0
        D09_ATH
        40.0
        0.0
        E09_ATH
        4.0
        0.0
        F09_ATH
        15.0
        0.0
        G09_ATH
        36.0
        0.0
        H09_ATH
        20.0
        0.0
        A10_ATH
        4210
        0.3
        B10_ATH
        5629
        0.4
        C10_ATH
        4098
        0.3
        D10_ATH
        6811
        0.4
        E10_ATH
        3493
        0.2
        F10_ATH
        2050
        0.1
        G10_ATH
        12495
        0.8
        H10_ATH
        6559
        0.4
        A11_ATH
        4825
        0.3
        B11_ATH
        3448
        0.2
        C11_ATH
        7781
        0.5
        D11_ATH
        6538
        0.4
        E11_ATH
        6036
        0.4
        F11_ATH
        6940
        0.4
        A01_CHI
        4768
        0.3
        B01_CHI
        4441
        0.3
        C01_CHI
        3148
        0.2
        D01_CHI
        2970
        0.2
        E01_CHI
        1113
        0.1
        F01_CHI
        2072
        0.1
        G01_CHI
        5625
        0.4
        H01_CHI
        3484
        0.2
        A02_CHI
        2990
        0.2
        B02_CHI
        1721
        0.1
        C02_CHI
        3311
        0.2
        D02_CHI
        1955
        0.1
        E02_CHI
        1521
        0.1
        F02_CHI
        1801
        0.1
        G02_CHI
        2144
        0.1
        H02_CHI
        3082
        0.2
        A03_CHI
        2305
        0.1
        B03_CHI
        2881
        0.2
        C03_CHI
        3844
        0.2
        D03_CHI
        2495
        0.2
        E03_CHI
        3575
        0.2
        F03_CHI
        502.0
        0.0
        G03_CHI
        4037
        0.3
        H03_CHI
        3540
        0.2
        A04_CHI
        1977
        0.1
        B04_CHI
        2659
        0.2
        C04_CHI
        2692
        0.2
        D04_CHI
        2396
        0.2
        E04_CHI
        495.0
        0.0
        F04_CHI
        410.0
        0.0
        G04_CHI
        3209
        0.2
        H04_CHI
        3354
        0.2
        A05_CHI
        3089
        0.2
        B05_CHI
        4200
        0.3
        C05_CHI
        5368
        0.3
        D05_CHI
        3753
        0.2
        E05_CHI
        2087
        0.1
        F05_CHI
        2908
        0.2
        G05_CHI
        5516
        0.4
        H05_CHI
        5560
        0.4
        A06_CHI
        4219
        0.3
        B06_CHI
        7558
        0.5
        C06_CHI
        14153
        0.9
        D06_CHI
        5131
        0.3
        E06_CHI
        2998
        0.2
        F06_CHI
        2678
        0.2
        G06_CHI
        7529
        0.5
        H06_CHI
        4985
        0.3
        A07_CHI
        10123
        0.6
        B07_CHI
        6486
        0.4
        C07_CHI
        10761
        0.7
        D07_CHI
        6530
        0.4
        E07_CHI
        5809
        0.4
        F07_CHI
        4414
        0.3
        G07_CHI
        9063
        0.6
        H07_CHI
        6984
        0.4
        A08_CHI
        2856
        0.2
        B08_CHI
        10685
        0.7
        C08_CHI
        5783
        0.4
        D08_CHI
        7808
        0.5
        E08_CHI
        7306
        0.5
        F08_CHI
        8750
        0.6
        G08_CHI
        12475
        0.8
        H08_CHI
        8309
        0.5
        A09_CHI
        8017
        0.5
        B09_CHI
        8480
        0.5
        C09_CHI
        8390
        0.5
        D09_CHI
        4758
        0.3
        E09_CHI
        4392
        0.3
        F09_CHI
        1021
        0.1
        G09_CHI
        8486
        0.5
        H09_CHI
        12885
        0.8
        A10_CHI
        6291
        0.4
        B10_CHI
        6951
        0.4
        C10_CHI
        4832
        0.3
        D10_CHI
        4918
        0.3
        E10_CHI
        3458
        0.2
        A11_CHI
        3845
        0.2
        B11_CHI
        4236
        0.3
        C11_CHI
        7076
        0.5
        D11_CHI
        10630
        0.7
        E11_CHI
        4014
        0.3
        A12_CHI
        4084
        0.3
        B12_CHI
        6058
        0.4
        C12_CHI
        4275
        0.3
        D12_CHI
        1554
        0.1
        E12_CHI
        3200
        0.2
        A01_DVE
        6907
        0.4
        B01_DVE
        12000
        0.8
        C01_DVE
        14269
        0.9
        D01_DVE
        9229
        0.6
        E01_DVE
        8082
        0.5
        F01_DVE
        11398
        0.7
        G01_DVE
        4122
        0.3
        H01_DVE
        12600
        0.8
        A02_DVE
        6753
        0.4
        B02_DVE
        10404
        0.7
        C02_DVE
        12385
        0.8
        D02_DVE
        3914
        0.2
        E02_DVE
        24681
        1.6
        F02_DVE
        8692
        0.6
        G02_DVE
        7293
        0.5
        H02_DVE
        14106
        0.9
        A03_DVE
        7902
        0.5
        B03_DVE
        12529
        0.8
        C03_DVE
        15571
        1.0
        D03_DVE
        10323
        0.7
        E03_DVE
        2223
        0.1
        F03_DVE
        11691
        0.7
        G03_DVE
        8282
        0.5
        H03_DVE
        13326
        0.8
        A04_DVE
        4332
        0.3
        B04_DVE
        11217
        0.7
        C04_DVE
        15434
        1.0
        D04_DVE
        11477
        0.7
        E04_DVE
        8696
        0.6
        F04_DVE
        9942
        0.6
        G04_DVE
        6550
        0.4
        H04_DVE
        11113
        0.7
        A05_DVE
        2267
        0.1
        B05_DVE
        3721
        0.2
        C05_DVE
        4154
        0.3
        D05_DVE
        1222
        0.1
        E05_DVE
        2761
        0.2
        F05_DVE
        3488
        0.2
        G05_DVE
        2757
        0.2
        H05_DVE
        3503
        0.2
        A06_DVE
        4139
        0.3
        B06_DVE
        5300
        0.3
        C06_DVE
        4865
        0.3
        D06_DVE
        3753
        0.2
        E06_DVE
        2529
        0.2
        F06_DVE
        8086
        0.5
        G06_DVE
        5081
        0.3
        A07_DVE
        6067
        0.4
        B07_DVE
        4342
        0.3
        C07_DVE
        4918
        0.3
        D07_DVE
        1624
        0.1
        E07_DVE
        3485
        0.2
        F07_DVE
        6199
        0.4
        G07_DVE
        2712
        0.2
        A08_DVE
        5620
        0.4
        B08_DVE
        9418
        0.6
        C08_DVE
        12612
        0.8
        D08_DVE
        9408
        0.6
        E08_DVE
        8422
        0.5
        F08_DVE
        10991
        0.7
        G08_DVE
        12385
        0.8
        A09_DVE
        9074
        0.6
        B09_DVE
        15147
        1.0
        C09_DVE
        26318
        1.7
        D09_DVE
        4140
        0.3
        E09_DVE
        540.0
        0.0
        F09_DVE
        7303
        0.5
        G09_DVE
        5265
        0.3
        A10_DVE
        3670
        0.2
        B10_DVE
        7163
        0.5
        C10_DVE
        6160
        0.4
        D10_DVE
        5688
        0.4
        E10_DVE
        4016
        0.3
        F10_DVE
        5874
        0.4
        G10_DVE
        6072
        0.4
        A11_DVE
        4935
        0.3
        B11_DVE
        5981
        0.4
        C11_DVE
        4889
        0.3
        D11_DVE
        6023
        0.4
        E11_DVE
        53.0
        0.0
        F11_DVE
        16557
        1.1
        G11_DVE
        13721
        0.9
        A12_DVE
        11.0
        0.0
        B12_DVE
        15002
        1.0
        C12_DVE
        6354
        0.4
        D12_DVE
        4501
        0.3
        E12_DVE
        7381
        0.5
        F12_DVE
        6572
        0.4
        G12_DVE
        5550
        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 (%)
        CATCGTGACTAGAGCT
        758.0
        0.7
        CATCGTGATCTTTCCC
        480.0
        0.5
        AGCATACCTCTTTCCC
        455.0
        0.4
        CGTCATACTCTTTCCC
        451.0
        0.4
        TCTAGACTTCTTTCCC
        447.0
        0.4
        CATCGTGACGTCGCTT
        430.0
        0.4
        TCAGTCTATCTTTCCC
        407.0
        0.4
        TCCTCATGTCTTTCCC
        386.0
        0.4
        ATAGCGCTTCTTTCCC
        384.0
        0.4
        AGCATACCCGTCGCTT
        357.0
        0.3
        CAGTAGGTCGTCGCTT
        315.0
        0.3
        ATAGCGCTCGTCGCTT
        309.0
        0.3
        AGCATACCACGTCTGT
        296.0
        0.3
        CGTCATACCGTCGCTT
        277.0
        0.3
        TCTAGACTCGTCGCTT
        258.0
        0.2
        CAGTAGGTTCTTTCCC
        257.0
        0.2
        ATGAGCTCTCTTTCCC
        251.0
        0.2
        TATAGCGAACGCGTGA
        251.0
        0.2
        CATCGTGAACGTCTGT
        227.0
        0.2
        TATAGCGACGATCTAC
        225.0
        0.2

        Lane Statistics

        Showing 1/1 rows and 4/4 columns.
        LaneTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        1.0
        1993811
        1568122
        6.8
        1.7

        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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