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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 2022-02-24, 01:33 based on data in: /scratch/gencore/logs/html/H35CCBGXL/merged


        General Statistics

        Showing 70/70 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        H35CCBGXL_n01_FebB2_1
        64.3%
        44%
        5.8
        H35CCBGXL_n01_FebB2_10
        37.3%
        43%
        6.1
        H35CCBGXL_n01_FebB2_11
        35.3%
        43%
        8.4
        H35CCBGXL_n01_FebB2_12
        38.3%
        43%
        6.1
        H35CCBGXL_n01_FebB2_13
        30.8%
        43%
        11.6
        H35CCBGXL_n01_FebB2_14
        53.4%
        44%
        11.1
        H35CCBGXL_n01_FebB2_15
        55.1%
        45%
        5.6
        H35CCBGXL_n01_FebB2_16
        52.4%
        44%
        7.4
        H35CCBGXL_n01_FebB2_17
        63.5%
        45%
        6.4
        H35CCBGXL_n01_FebB2_18
        64.4%
        44%
        6.6
        H35CCBGXL_n01_FebB2_19
        39.0%
        43%
        7.6
        H35CCBGXL_n01_FebB2_2
        76.1%
        47%
        7.7
        H35CCBGXL_n01_FebB2_20
        71.2%
        44%
        10.2
        H35CCBGXL_n01_FebB2_21
        77.3%
        44%
        4.5
        H35CCBGXL_n01_FebB2_22
        62.9%
        45%
        5.8
        H35CCBGXL_n01_FebB2_23
        65.3%
        44%
        5.9
        H35CCBGXL_n01_FebB2_24
        53.6%
        46%
        6.6
        H35CCBGXL_n01_FebB2_3
        62.5%
        44%
        6.4
        H35CCBGXL_n01_FebB2_4
        44.4%
        44%
        8.0
        H35CCBGXL_n01_FebB2_5
        46.6%
        43%
        6.9
        H35CCBGXL_n01_FebB2_6
        64.4%
        44%
        7.0
        H35CCBGXL_n01_FebB2_7
        66.8%
        44%
        6.2
        H35CCBGXL_n01_FebB2_8
        39.0%
        44%
        8.6
        H35CCBGXL_n01_FebB2_9
        57.2%
        44%
        7.0
        H35CCBGXL_n01_FebB3_1
        31.0%
        43%
        7.0
        H35CCBGXL_n01_FebB3_10
        30.8%
        43%
        7.4
        H35CCBGXL_n01_FebB3_11
        26.5%
        42%
        8.0
        H35CCBGXL_n01_FebB3_12
        48.0%
        45%
        6.4
        H35CCBGXL_n01_FebB3_13
        75.1%
        45%
        8.3
        H35CCBGXL_n01_FebB3_14
        33.7%
        45%
        6.9
        H35CCBGXL_n01_FebB3_15
        29.9%
        44%
        6.2
        H35CCBGXL_n01_FebB3_16
        29.2%
        43%
        6.6
        H35CCBGXL_n01_FebB3_17
        33.2%
        43%
        6.8
        H35CCBGXL_n01_FebB3_18
        47.0%
        44%
        6.4
        H35CCBGXL_n01_FebB3_19
        33.5%
        44%
        6.8
        H35CCBGXL_n01_FebB3_2
        29.4%
        43%
        8.0
        H35CCBGXL_n01_FebB3_20
        47.3%
        43%
        6.5
        H35CCBGXL_n01_FebB3_21
        43.1%
        44%
        5.9
        H35CCBGXL_n01_FebB3_22
        27.1%
        42%
        6.6
        H35CCBGXL_n01_FebB3_23
        65.7%
        46%
        4.0
        H35CCBGXL_n01_FebB3_24
        62.0%
        44%
        4.7
        H35CCBGXL_n01_FebB3_3
        41.8%
        45%
        6.3
        H35CCBGXL_n01_FebB3_4
        32.2%
        44%
        7.6
        H35CCBGXL_n01_FebB3_5
        43.8%
        44%
        6.9
        H35CCBGXL_n01_FebB3_6
        29.4%
        43%
        6.6
        H35CCBGXL_n01_FebB3_7
        81.6%
        45%
        2.5
        H35CCBGXL_n01_FebB3_8
        48.7%
        45%
        11.4
        H35CCBGXL_n01_FebB3_9
        32.4%
        44%
        7.6
        H35CCBGXL_n01_FebB4_1
        26.0%
        37%
        7.4
        H35CCBGXL_n01_FebB4_10
        27.8%
        39%
        6.7
        H35CCBGXL_n01_FebB4_11
        36.3%
        41%
        8.1
        H35CCBGXL_n01_FebB4_12
        28.0%
        40%
        7.3
        H35CCBGXL_n01_FebB4_13
        30.1%
        41%
        6.8
        H35CCBGXL_n01_FebB4_14
        31.9%
        41%
        6.1
        H35CCBGXL_n01_FebB4_15
        21.3%
        39%
        6.7
        H35CCBGXL_n01_FebB4_16
        33.8%
        41%
        6.4
        H35CCBGXL_n01_FebB4_17
        27.3%
        40%
        6.7
        H35CCBGXL_n01_FebB4_18
        26.8%
        39%
        6.8
        H35CCBGXL_n01_FebB4_19
        32.3%
        40%
        6.9
        H35CCBGXL_n01_FebB4_2
        24.2%
        40%
        6.8
        H35CCBGXL_n01_FebB4_20
        30.9%
        40%
        6.1
        H35CCBGXL_n01_FebB4_21
        29.2%
        40%
        6.6
        H35CCBGXL_n01_FebB4_3
        33.9%
        41%
        6.4
        H35CCBGXL_n01_FebB4_4
        22.9%
        39%
        6.3
        H35CCBGXL_n01_FebB4_5
        19.6%
        38%
        6.3
        H35CCBGXL_n01_FebB4_6
        24.7%
        39%
        5.9
        H35CCBGXL_n01_FebB4_7
        20.5%
        39%
        6.5
        H35CCBGXL_n01_FebB4_8
        19.0%
        38%
        6.3
        H35CCBGXL_n01_FebB4_9
        26.9%
        40%
        6.9
        H35CCBGXL_n01_undetermined
        87.1%
        44%
        82.7

        Barcodes of Undetermined Reads


        We have determined the barcodes of your undetermined reads (reads containing a barcode that you did not encode in your metadata). Here are the top 20 barcodes belonging to the undetermined reads. The full list is available here.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGG
        76969416.0
        93.1
        NNNNNN
        242446.0
        0.3
        GGGGGC
        100513.0
        0.1
        GCGGGG
        95231.0
        0.1
        GGGGCG
        92350.0
        0.1
        GGGCGG
        77105.0
        0.1
        GGCGGG
        67164.0
        0.1
        GGGGTG
        62075.0
        0.1
        GTGGGG
        56301.0
        0.1
        GGGTGG
        55036.0
        0.1
        TGGGGG
        51075.0
        0.1
        CGGGGG
        51012.0
        0.1
        TTATGA
        39366.0
        0.1
        CAATTA
        27168.0
        0.0
        GAAGGA
        26675.0
        0.0
        GTGAGA
        24881.0
        0.0
        TGGTAA
        24714.0
        0.0
        TGATTA
        24609.0
        0.0
        AAAAAA
        24390.0
        0.0
        CGAGGA
        24239.0
        0.0

        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 70/70 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        82689442
        14.8
        FebB2_1
        5838772
        1.0
        FebB2_2
        7657969
        1.4
        FebB2_3
        6424125
        1.2
        FebB2_4
        7982903
        1.4
        FebB2_5
        6924723
        1.2
        FebB2_6
        7007126
        1.3
        FebB2_7
        6224777
        1.1
        FebB2_8
        8640899
        1.6
        FebB2_9
        6994185
        1.3
        FebB2_10
        6073663
        1.1
        FebB2_11
        8405061
        1.5
        FebB2_12
        6136583
        1.1
        FebB2_13
        11617786
        2.1
        FebB2_14
        11137417
        2.0
        FebB2_15
        5614279
        1.0
        FebB2_16
        7436512
        1.3
        FebB2_17
        6407179
        1.1
        FebB2_18
        6566533
        1.2
        FebB2_19
        7638976
        1.4
        FebB2_20
        10230275
        1.8
        FebB2_21
        4451047
        0.8
        FebB2_22
        5798624
        1.0
        FebB2_23
        5851565
        1.1
        FebB2_24
        6644025
        1.2
        FebB3_1
        7028915
        1.3
        FebB3_2
        7960965
        1.4
        FebB3_3
        6308611
        1.1
        FebB3_4
        7580927
        1.4
        FebB3_5
        6868754
        1.2
        FebB3_6
        6618189
        1.2
        FebB3_7
        2488378
        0.4
        FebB3_8
        11376076
        2.0
        FebB3_9
        7600914
        1.4
        FebB3_10
        7355530
        1.3
        FebB3_11
        7999042
        1.4
        FebB3_12
        6424341
        1.2
        FebB3_13
        8339230
        1.5
        FebB3_14
        6905152
        1.2
        FebB3_15
        6241074
        1.1
        FebB3_16
        6646633
        1.2
        FebB3_17
        6775162
        1.2
        FebB3_18
        6370226
        1.1
        FebB3_19
        6833229
        1.2
        FebB3_20
        6518263
        1.2
        FebB3_21
        5864229
        1.1
        FebB3_22
        6607582
        1.2
        FebB3_23
        4027340
        0.7
        FebB3_24
        4733581
        0.8
        FebB4_1
        7415486
        1.3
        FebB4_2
        6774023
        1.2
        FebB4_3
        6357702
        1.1
        FebB4_4
        6323521
        1.1
        FebB4_5
        6303228
        1.1
        FebB4_6
        5902994
        1.1
        FebB4_7
        6470499
        1.2
        FebB4_8
        6260822
        1.1
        FebB4_9
        6944397
        1.2
        FebB4_10
        6691728
        1.2
        FebB4_11
        8073541
        1.4
        FebB4_12
        7330397
        1.3
        FebB4_13
        6784513
        1.2
        FebB4_14
        6052719
        1.1
        FebB4_15
        6659665
        1.2
        FebB4_16
        6376479
        1.1
        FebB4_17
        6711634
        1.2
        FebB4_18
        6814098
        1.2
        FebB4_19
        6910297
        1.2
        FebB4_20
        6114075
        1.1
        FebB4_21
        6622920
        1.2

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        4.0
        658684368
        557761527
        14.8
        13.1

        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 (76bp).

        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..