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        Download the raw data used to create the plots in this report below:

        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 2020-11-04, 11:41 based on data in: /mnt/bfx6/bfx/adam_trewin/set2


        General Statistics

        Showing 132/132 rows and 6/8 columns.
        Sample Name% AlignedM Aligned% Trimmed% Dups% GCM Seqs
        10_S10_R1_001
        89.7%
        36.2
        1.7%
        72.7%
        48%
        40.3
        10_S10_R2_001
        37.1%
        49%
        40.3
        11_S11_R1_001
        90.7%
        31.6
        1.7%
        72.0%
        48%
        34.9
        11_S11_R2_001
        35.2%
        49%
        34.9
        12_S12_R1_001
        88.9%
        34.9
        1.7%
        71.2%
        49%
        39.3
        12_S12_R2_001
        37.8%
        51%
        39.3
        13_S13_R1_001
        89.6%
        34.6
        1.7%
        70.2%
        49%
        38.6
        13_S13_R2_001
        36.1%
        50%
        38.6
        14_S14_R1_001
        90.5%
        31.8
        1.7%
        70.8%
        48%
        35.2
        14_S14_R2_001
        35.7%
        49%
        35.2
        15_S15_R1_001
        90.1%
        29.2
        1.7%
        69.5%
        48%
        32.4
        15_S15_R2_001
        34.8%
        49%
        32.4
        16_S16_R1_001
        90.5%
        32.2
        1.7%
        70.0%
        48%
        35.6
        16_S16_R2_001
        33.8%
        49%
        35.6
        17_S17_R1_001
        88.2%
        35.5
        1.7%
        80.5%
        48%
        40.2
        17_S17_R2_001
        55.9%
        50%
        40.2
        18_S18_R1_001
        89.9%
        40.0
        1.8%
        75.6%
        47%
        44.5
        18_S18_R2_001
        45.8%
        49%
        44.5
        19_S19_R1_001
        89.9%
        36.5
        1.7%
        77.7%
        47%
        40.6
        19_S19_R2_001
        48.5%
        50%
        40.6
        1_S1_R1_001
        90.4%
        26.7
        1.8%
        73.1%
        48%
        29.5
        1_S1_R2_001
        45.1%
        50%
        29.5
        20_S20_R1_001
        88.5%
        35.0
        1.7%
        79.4%
        48%
        39.5
        20_S20_R2_001
        52.9%
        50%
        39.5
        21_S21_R1_001
        88.0%
        46.8
        1.7%
        80.5%
        48%
        53.2
        21_S21_R2_001
        54.7%
        50%
        53.2
        22_S22_R1_001
        89.4%
        33.2
        1.8%
        79.8%
        47%
        37.1
        22_S22_R2_001
        56.2%
        50%
        37.1
        23_S23_R1_001
        89.2%
        36.7
        1.7%
        78.8%
        48%
        41.1
        23_S23_R2_001
        49.9%
        50%
        41.1
        24_S24_R1_001
        88.7%
        35.5
        1.8%
        72.3%
        49%
        40.0
        24_S24_R2_001
        43.0%
        51%
        40.0
        25_S25_R1_001
        87.6%
        40.9
        1.7%
        80.6%
        48%
        46.7
        25_S25_R2_001
        56.2%
        51%
        46.7
        26_S26_R1_001
        78.9%
        34.3
        1.8%
        76.0%
        50%
        43.4
        26_S26_R2_001
        47.6%
        52%
        43.4
        27_S27_R1_001
        88.6%
        34.8
        1.7%
        75.1%
        48%
        39.2
        27_S27_R2_001
        45.4%
        50%
        39.2
        28_S28_R1_001
        81.5%
        36.2
        1.7%
        75.4%
        50%
        44.4
        28_S28_R2_001
        45.5%
        52%
        44.4
        29_S29_R1_001
        82.7%
        27.2
        1.7%
        78.1%
        49%
        32.9
        29_S29_R2_001
        51.5%
        51%
        32.9
        2_S2_R1_001
        90.1%
        21.5
        1.7%
        74.4%
        48%
        23.8
        2_S2_R2_001
        44.7%
        50%
        23.8
        30_S30_R1_001
        87.1%
        32.1
        1.8%
        73.3%
        49%
        36.8
        30_S30_R2_001
        43.0%
        51%
        36.8
        31_S31_R1_001
        87.3%
        31.3
        1.7%
        74.8%
        48%
        35.8
        31_S31_R2_001
        44.8%
        50%
        35.8
        32_S32_R1_001
        89.0%
        25.9
        1.7%
        66.2%
        48%
        29.1
        32_S32_R2_001
        32.5%
        49%
        29.1
        33_S33_R1_001
        83.9%
        23.5
        2.5%
        33.2%
        43%
        28.0
        33_S33_R2_001
        21.1%
        45%
        28.0
        34_S34_R1_001
        84.5%
        22.9
        2.5%
        65.6%
        42%
        27.1
        34_S34_R2_001
        51.8%
        44%
        27.1
        35_S35_R1_001
        83.3%
        22.0
        2.8%
        15.7%
        45%
        26.5
        35_S35_R2_001
        10.2%
        47%
        26.5
        36_S36_R1_001
        83.6%
        15.7
        2.7%
        12.6%
        45%
        18.8
        36_S36_R2_001
        6.7%
        47%
        18.8
        37_S37_R1_001
        83.7%
        20.6
        2.5%
        16.3%
        45%
        24.7
        37_S37_R2_001
        9.6%
        47%
        24.7
        38_S38_R1_001
        83.3%
        21.5
        2.6%
        12.7%
        45%
        25.8
        38_S38_R2_001
        7.3%
        47%
        25.8
        39_S39_R1_001
        83.3%
        16.1
        2.7%
        17.9%
        45%
        19.4
        39_S39_R2_001
        9.7%
        46%
        19.4
        3_S3_R1_001
        89.7%
        26.6
        1.8%
        72.8%
        48%
        29.7
        3_S3_R2_001
        41.3%
        50%
        29.7
        40_S40_R1_001
        83.5%
        14.8
        2.6%
        9.7%
        45%
        17.7
        40_S40_R2_001
        5.0%
        47%
        17.7
        41_S41_R1_001
        82.8%
        13.7
        2.9%
        9.4%
        45%
        16.5
        41_S41_R2_001
        5.5%
        46%
        16.5
        42_S42_R1_001
        83.2%
        12.0
        2.8%
        8.5%
        45%
        14.5
        42_S42_R2_001
        4.6%
        47%
        14.5
        43_S43_R1_001
        83.5%
        26.6
        2.9%
        19.1%
        45%
        31.9
        43_S43_R2_001
        13.7%
        46%
        31.9
        44_S44_R1_001
        83.4%
        16.8
        3.0%
        10.2%
        45%
        20.2
        44_S44_R2_001
        5.8%
        46%
        20.2
        45_S45_R1_001
        83.5%
        19.8
        2.6%
        11.4%
        45%
        23.7
        45_S45_R2_001
        5.6%
        46%
        23.7
        46_S46_R1_001
        83.5%
        17.7
        2.9%
        13.0%
        45%
        21.2
        46_S46_R2_001
        7.1%
        46%
        21.2
        47_S47_R1_001
        82.3%
        24.4
        2.6%
        53.5%
        44%
        29.7
        47_S47_R2_001
        39.2%
        45%
        29.7
        48_S48_R1_001
        83.5%
        15.9
        2.8%
        13.8%
        45%
        19.0
        48_S48_R2_001
        7.3%
        46%
        19.0
        49_S49_R1_001
        83.9%
        20.2
        2.6%
        13.0%
        45%
        24.1
        49_S49_R2_001
        7.5%
        46%
        24.1
        4_S4_R1_001
        91.5%
        30.5
        1.7%
        68.9%
        48%
        33.4
        4_S4_R2_001
        35.0%
        50%
        33.4
        50_S50_R1_001
        83.5%
        25.3
        2.8%
        15.9%
        45%
        30.3
        50_S50_R2_001
        10.2%
        46%
        30.3
        51_S51_R1_001
        82.8%
        30.9
        2.7%
        16.1%
        45%
        37.4
        51_S51_R2_001
        9.7%
        46%
        37.4
        52_S52_R1_001
        81.3%
        27.6
        3.5%
        40.5%
        44%
        34.0
        52_S52_R2_001
        32.9%
        45%
        34.0
        53_S53_R1_001
        81.7%
        24.9
        3.3%
        23.3%
        44%
        30.6
        53_S53_R2_001
        16.5%
        45%
        30.6
        54_S54_R1_001
        83.7%
        22.9
        2.8%
        13.5%
        45%
        27.4
        54_S54_R2_001
        7.8%
        46%
        27.4
        55_S55_R1_001
        84.2%
        21.9
        2.7%
        10.6%
        45%
        26.0
        55_S55_R2_001
        6.3%
        46%
        26.0
        56_S56_R1_001
        81.6%
        28.4
        3.4%
        29.4%
        43%
        34.8
        56_S56_R2_001
        21.9%
        45%
        34.8
        57_S57_R1_001
        83.4%
        24.9
        2.8%
        17.9%
        44%
        29.8
        57_S57_R2_001
        10.9%
        45%
        29.8
        58_S58_R1_001
        83.4%
        26.4
        2.8%
        27.7%
        44%
        31.6
        58_S58_R2_001
        18.3%
        45%
        31.6
        59_S59_R1_001
        81.6%
        27.1
        2.8%
        47.9%
        43%
        33.2
        59_S59_R2_001
        35.3%
        44%
        33.2
        5_S5_R1_001
        89.0%
        33.9
        1.7%
        73.7%
        48%
        38.1
        5_S5_R2_001
        41.1%
        50%
        38.1
        60_S60_R1_001
        82.4%
        28.8
        3.4%
        48.0%
        43%
        35.0
        60_S60_R2_001
        37.8%
        44%
        35.0
        61_S61_R1_001
        82.7%
        25.8
        2.8%
        19.5%
        45%
        31.2
        61_S61_R2_001
        13.0%
        46%
        31.2
        62_S62_R1_001
        83.0%
        26.1
        2.8%
        18.0%
        45%
        31.4
        62_S62_R2_001
        12.1%
        46%
        31.4
        63_S63_R1_001
        83.1%
        24.6
        2.9%
        19.3%
        45%
        29.6
        63_S63_R2_001
        13.1%
        46%
        29.6
        64_S64_R1_001
        83.0%
        26.0
        3.0%
        36.1%
        44%
        31.4
        64_S64_R2_001
        26.8%
        45%
        31.4
        6_S6_R1_001
        90.3%
        30.0
        1.7%
        71.8%
        48%
        33.3
        6_S6_R2_001
        39.2%
        50%
        33.3
        7_S7_R1_001
        90.3%
        29.5
        1.7%
        69.9%
        48%
        32.6
        7_S7_R2_001
        34.9%
        50%
        32.6
        8_S8_R1_001
        89.7%
        27.6
        1.8%
        65.6%
        49%
        30.8
        8_S8_R2_001
        33.3%
        50%
        30.8
        95_S95_R1_001
        15.7%
        5.8
        1.9%
        96.4%
        45%
        37.0
        95_S95_R2_001
        76.9%
        46%
        37.0
        96_S96_R1_001
        84.0%
        30.2
        3.0%
        94.7%
        40%
        36.0
        96_S96_R2_001
        78.2%
        41%
        36.0
        9_S9_R1_001
        90.3%
        28.0
        1.8%
        70.2%
        47%
        31.0
        9_S9_R2_001
        33.3%
        49%
        31.0

        STAR

        STAR is an ultrafast universal RNA-seq aligner.

        Alignment Scores

        loading..

        Gene Counts

        Statistics from results generated using --quantMode GeneCounts. The three tabs show counts for unstranded RNA-seq, counts for the 1st read strand aligned with RNA and counts for the 2nd read strand aligned with RNA.

           
        loading..

        Skewer

        Skewer is an adapter trimming tool specially designed for processing next-generation sequencing (NGS) paired-end sequences.

        loading..

        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).


        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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