Multiqc for JA21499 SA22026

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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 1.6

        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

        Multiqc for JA21499 SA22026

        Multiqc report for Fastq files in job JA21499, run SA22026

        Report generated on 2022-02-15, 15:48 based on data in: /raida/gsafdata/jobinfo/qc/JA21499/SA22026/fastqc


        General Statistics

        Showing 144/144 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        D54_S145_L001_R1_001
        16.0%
        38%
        2.3
        D54_S145_L002_R1_001
        16.7%
        38%
        2.3
        D55_S146_L001_R1_001
        11.5%
        36%
        3.1
        D55_S146_L002_R1_001
        12.2%
        36%
        3.2
        D56_S136_L001_R1_001
        15.3%
        37%
        3.1
        D56_S136_L002_R1_001
        15.9%
        37%
        3.2
        D57_S143_L001_R1_001
        18.5%
        38%
        2.8
        D57_S143_L002_R1_001
        19.1%
        38%
        2.8
        D58_S144_L001_R1_001
        11.3%
        36%
        4.1
        D58_S144_L002_R1_001
        12.0%
        36%
        4.2
        D59_S142_L001_R1_001
        14.8%
        38%
        2.8
        D59_S142_L002_R1_001
        15.4%
        38%
        2.8
        M41_S161_L001_R1_001
        15.2%
        37%
        2.8
        M41_S161_L002_R1_001
        15.9%
        37%
        2.9
        M42_S169_L001_R1_001
        16.6%
        38%
        2.7
        M42_S169_L002_R1_001
        17.3%
        38%
        2.7
        M43_S160_L001_R1_001
        21.9%
        39%
        2.9
        M43_S160_L002_R1_001
        22.5%
        39%
        2.9
        M44_S163_L001_R1_001
        16.1%
        37%
        2.2
        M44_S163_L002_R1_001
        16.7%
        37%
        2.3
        M45_S140_L001_R1_001
        15.5%
        37%
        2.9
        M45_S140_L002_R1_001
        16.2%
        37%
        2.9
        M46_S141_L001_R1_001
        16.8%
        38%
        2.8
        M46_S141_L002_R1_001
        17.6%
        38%
        2.8
        M47_S162_L001_R1_001
        17.0%
        38%
        3.1
        M47_S162_L002_R1_001
        17.8%
        38%
        3.1
        M48_S137_L001_R1_001
        16.0%
        37%
        2.5
        M48_S137_L002_R1_001
        16.5%
        37%
        2.6
        M49_S139_L001_R1_001
        17.3%
        38%
        3.4
        M49_S139_L002_R1_001
        18.1%
        38%
        3.5
        M89_S138_L001_R1_001
        12.1%
        36%
        3.5
        M89_S138_L002_R1_001
        12.9%
        36%
        3.5
        M90_S147_L001_R1_001
        16.2%
        38%
        2.4
        M90_S147_L002_R1_001
        16.7%
        38%
        2.4
        N41_S164_L001_R1_001
        16.3%
        37%
        2.7
        N41_S164_L002_R1_001
        17.0%
        37%
        2.7
        N42_S167_L001_R1_001
        14.6%
        37%
        2.6
        N42_S167_L002_R1_001
        15.2%
        37%
        2.7
        N43_S165_L001_R1_001
        15.4%
        37%
        3.1
        N43_S165_L002_R1_001
        16.2%
        37%
        3.2
        N44_S171_L001_R1_001
        16.9%
        37%
        2.7
        N44_S171_L002_R1_001
        17.8%
        37%
        2.8
        N45_S166_L001_R1_001
        18.5%
        38%
        2.9
        N45_S166_L002_R1_001
        19.1%
        38%
        3.0
        N46_S170_L001_R1_001
        17.1%
        37%
        2.9
        N46_S170_L002_R1_001
        17.8%
        37%
        2.9
        N47_S168_L001_R1_001
        16.3%
        39%
        2.7
        N47_S168_L002_R1_001
        17.1%
        39%
        2.7
        N48_S194_L001_R1_001
        13.4%
        37%
        2.2
        N48_S194_L002_R1_001
        14.1%
        37%
        2.2
        N49_S185_L001_R1_001
        13.3%
        37%
        2.5
        N49_S185_L002_R1_001
        13.7%
        37%
        2.5
        N50_S187_L001_R1_001
        18.0%
        38%
        3.1
        N50_S187_L002_R1_001
        18.4%
        38%
        3.1
        N51_S186_L001_R1_001
        17.3%
        39%
        2.6
        N51_S186_L002_R1_001
        18.0%
        39%
        2.7
        N52_S184_L001_R1_001
        12.1%
        36%
        3.7
        N52_S184_L002_R1_001
        12.8%
        36%
        3.8
        N53_S188_L001_R1_001
        17.9%
        38%
        2.9
        N53_S188_L002_R1_001
        18.6%
        38%
        3.0
        N54_S193_L001_R1_001
        21.1%
        38%
        3.3
        N54_S193_L002_R1_001
        21.6%
        38%
        3.3
        N55_S190_L001_R1_001
        15.7%
        37%
        2.1
        N55_S190_L002_R1_001
        16.4%
        37%
        2.1
        N56_S192_L001_R1_001
        36.2%
        37%
        0.5
        N56_S192_L002_R1_001
        36.9%
        37%
        0.5
        N57_S191_L001_R1_001
        10.0%
        35%
        3.0
        N57_S191_L002_R1_001
        10.6%
        35%
        3.0
        N58_S195_L001_R1_001
        19.0%
        39%
        2.8
        N58_S195_L002_R1_001
        19.6%
        39%
        2.9
        N59_S189_L001_R1_001
        19.3%
        36%
        2.9
        N59_S189_L002_R1_001
        19.9%
        36%
        3.0
        R41_S181_L001_R1_001
        14.9%
        36%
        3.3
        R41_S181_L002_R1_001
        15.5%
        36%
        3.4
        R42_S175_L001_R1_001
        16.3%
        38%
        2.6
        R42_S175_L002_R1_001
        16.9%
        38%
        2.6
        R43_S177_L001_R1_001
        15.3%
        37%
        3.0
        R43_S177_L002_R1_001
        15.8%
        37%
        3.1
        R46_S178_L001_R1_001
        14.4%
        36%
        3.6
        R46_S178_L002_R1_001
        15.0%
        36%
        3.6
        R47_S179_L001_R1_001
        14.7%
        37%
        2.5
        R47_S179_L002_R1_001
        15.6%
        37%
        2.5
        R48_S176_L001_R1_001
        9.8%
        36%
        2.6
        R48_S176_L002_R1_001
        10.4%
        36%
        2.6
        R50_S174_L001_R1_001
        16.2%
        36%
        2.6
        R50_S174_L002_R1_001
        16.6%
        36%
        2.7
        R51_S205_L001_R1_001
        13.0%
        37%
        2.8
        R51_S205_L002_R1_001
        13.7%
        37%
        2.9
        R52_S204_L001_R1_001
        12.7%
        38%
        2.4
        R52_S204_L002_R1_001
        13.2%
        38%
        2.4
        R53_S199_L001_R1_001
        18.0%
        36%
        2.8
        R53_S199_L002_R1_001
        18.6%
        36%
        2.9
        R54_S206_L001_R1_001
        16.0%
        38%
        3.1
        R54_S206_L002_R1_001
        16.6%
        38%
        3.1
        R55_S196_L001_R1_001
        17.9%
        37%
        2.8
        R55_S196_L002_R1_001
        18.4%
        37%
        2.8
        R56_S197_L001_R1_001
        16.4%
        38%
        2.6
        R56_S197_L002_R1_001
        17.1%
        38%
        2.7
        R57_S200_L001_R1_001
        12.7%
        37%
        3.0
        R57_S200_L002_R1_001
        13.6%
        37%
        3.0
        R58_S198_L001_R1_001
        15.5%
        38%
        2.8
        R58_S198_L002_R1_001
        16.2%
        38%
        2.9
        R59_S202_L001_R1_001
        13.7%
        38%
        2.0
        R59_S202_L002_R1_001
        14.1%
        38%
        2.0
        R60_S201_L001_R1_001
        18.4%
        40%
        2.6
        R60_S201_L002_R1_001
        19.0%
        40%
        2.6
        R61_S207_L001_R1_001
        13.4%
        37%
        2.2
        R61_S207_L002_R1_001
        13.9%
        37%
        2.3
        R62_S203_L001_R1_001
        10.1%
        36%
        1.5
        R62_S203_L002_R1_001
        10.7%
        36%
        1.5
        R78_S180_L001_R1_001
        16.0%
        37%
        2.9
        R78_S180_L002_R1_001
        16.6%
        37%
        2.9
        T55_S158_L001_R1_001
        13.8%
        38%
        2.8
        T55_S158_L002_R1_001
        14.5%
        38%
        2.8
        T56_S155_L001_R1_001
        13.7%
        37%
        2.7
        T56_S155_L002_R1_001
        14.1%
        37%
        2.8
        T57_S157_L001_R1_001
        18.0%
        38%
        3.5
        T57_S157_L002_R1_001
        18.6%
        38%
        3.6
        T58_S152_L001_R1_001
        18.6%
        36%
        2.2
        T58_S152_L002_R1_001
        19.2%
        36%
        2.2
        T59_S151_L001_R1_001
        16.6%
        37%
        2.2
        T59_S151_L002_R1_001
        17.4%
        37%
        2.2
        T60_S149_L001_R1_001
        12.2%
        36%
        2.7
        T60_S149_L002_R1_001
        12.9%
        36%
        2.8
        T61_S156_L001_R1_001
        13.4%
        36%
        3.5
        T61_S156_L002_R1_001
        14.1%
        36%
        3.6
        T62_S148_L001_R1_001
        23.5%
        37%
        2.2
        T62_S148_L002_R1_001
        23.9%
        37%
        2.2
        X41_S183_L001_R1_001
        15.3%
        36%
        2.8
        X41_S183_L002_R1_001
        15.8%
        36%
        2.9
        X42_S182_L001_R1_001
        29.3%
        36%
        1.9
        X42_S182_L002_R1_001
        29.8%
        36%
        2.0
        X43_S173_L001_R1_001
        12.4%
        37%
        2.5
        X43_S173_L002_R1_001
        12.7%
        37%
        2.5
        X44_S172_L001_R1_001
        50.1%
        37%
        0.5
        X44_S172_L002_R1_001
        50.7%
        37%
        0.6
        X45_S159_L001_R1_001
        16.0%
        38%
        2.9
        X45_S159_L002_R1_001
        16.8%
        38%
        3.0
        X46_S153_L001_R1_001
        13.9%
        37%
        3.3
        X46_S153_L002_R1_001
        14.6%
        37%
        3.4
        X47_S150_L001_R1_001
        14.2%
        36%
        3.4
        X47_S150_L002_R1_001
        14.8%
        36%
        3.5
        X48_S154_L001_R1_001
        15.5%
        38%
        2.5
        X48_S154_L002_R1_001
        16.3%
        38%
        2.6

        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 (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.

        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).