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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 2020-12-22, 17:53 based on data in: /scratch/gencore/logs/html/000000000-JCJPN/1


        General Statistics

        Showing 96/96 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        000000000-JCJPN_l01_n01_HRB0060
        0.7%
        41%
        0.6
        000000000-JCJPN_l01_n01_HRB0062
        1.9%
        42%
        0.7
        000000000-JCJPN_l01_n01_HRB0069
        1.1%
        43%
        0.6
        000000000-JCJPN_l01_n01_HRB0070
        0.7%
        42%
        0.5
        000000000-JCJPN_l01_n01_HRB0071
        0.5%
        42%
        0.6
        000000000-JCJPN_l01_n01_HRB0084
        1.6%
        45%
        0.6
        000000000-JCJPN_l01_n01_HRB0092
        0.1%
        49%
        0.1
        000000000-JCJPN_l01_n01_HRB0093
        1.1%
        39%
        0.6
        000000000-JCJPN_l01_n01_HRB0094
        1.2%
        42%
        0.8
        000000000-JCJPN_l01_n01_HRB0171
        1.4%
        44%
        0.6
        000000000-JCJPN_l01_n01_HRB0172
        1.2%
        44%
        0.9
        000000000-JCJPN_l01_n01_HRB0192
        0.7%
        39%
        0.5
        000000000-JCJPN_l01_n01_HRB0203
        0.7%
        44%
        0.6
        000000000-JCJPN_l01_n01_HRB0204
        0.6%
        45%
        0.7
        000000000-JCJPN_l01_n01_HRB0206
        9.4%
        44%
        0.1
        000000000-JCJPN_l01_n01_HRB0208
        0.7%
        43%
        0.5
        000000000-JCJPN_l01_n01_HRB0209
        0.6%
        42%
        0.7
        000000000-JCJPN_l01_n01_HRB0210
        0.3%
        40%
        0.4
        000000000-JCJPN_l01_n01_HRB0211
        0.8%
        44%
        0.4
        000000000-JCJPN_l01_n01_HRB0249
        1.2%
        44%
        0.7
        000000000-JCJPN_l01_n01_HRB0250
        1.9%
        43%
        0.8
        000000000-JCJPN_l01_n01_HRB0256
        0.5%
        43%
        0.6
        000000000-JCJPN_l01_n01_HRB0257
        0.8%
        41%
        0.7
        000000000-JCJPN_l01_n01_HRB0258L
        0.5%
        44%
        0.3
        000000000-JCJPN_l01_n01_HRB0258R
        0.2%
        46%
        0.3
        000000000-JCJPN_l01_n01_HRB0259L
        0.6%
        42%
        0.3
        000000000-JCJPN_l01_n01_HRB0259R
        0.1%
        51%
        0.3
        000000000-JCJPN_l01_n01_HRB0260L
        0.3%
        41%
        0.3
        000000000-JCJPN_l01_n01_HRB0260R
        0.2%
        47%
        0.4
        000000000-JCJPN_l01_n01_HRB0261L
        0.4%
        40%
        0.3
        000000000-JCJPN_l01_n01_HRB0261R
        0.3%
        44%
        0.4
        000000000-JCJPN_l01_n01_HRB0263L
        0.5%
        42%
        0.5
        000000000-JCJPN_l01_n01_HRB0263R
        0.1%
        47%
        0.5
        000000000-JCJPN_l01_n01_HRB0268
        0.4%
        41%
        0.3
        000000000-JCJPN_l01_n01_HRB0269
        4.2%
        38%
        0.4
        000000000-JCJPN_l01_n01_HRB0270
        1.6%
        40%
        0.3
        000000000-JCJPN_l01_n01_HRB0271
        0.4%
        41%
        0.3
        000000000-JCJPN_l01_n01_HRB0272
        0.6%
        38%
        0.7
        000000000-JCJPN_l01_n01_HRB0273
        1.0%
        40%
        0.1
        000000000-JCJPN_l01_n01_HRBB1EB
        7.1%
        43%
        0.0
        000000000-JCJPN_l01_n01_HRBB1LB
        77.6%
        41%
        0.0
        000000000-JCJPN_l01_n01_HRBB2EB
        9.8%
        44%
        0.0
        000000000-JCJPN_l01_n01_HRBB2LB
        81.8%
        41%
        0.0
        000000000-JCJPN_l01_n01_HRBB3EB
        14.3%
        46%
        0.0
        000000000-JCJPN_l01_n01_HRBB3LB
        81.1%
        41%
        0.0
        000000000-JCJPN_l01_n01_HRBB4EB
        1.8%
        47%
        0.0
        000000000-JCJPN_l01_n01_HRBB4LB
        56.5%
        43%
        0.0
        000000000-JCJPN_l01_n01_undetermined
        62.4%
        44%
        0.7
        000000000-JCJPN_l01_n02_HRB0060
        0.7%
        41%
        0.6
        000000000-JCJPN_l01_n02_HRB0062
        1.8%
        43%
        0.7
        000000000-JCJPN_l01_n02_HRB0069
        1.0%
        42%
        0.6
        000000000-JCJPN_l01_n02_HRB0070
        0.7%
        43%
        0.5
        000000000-JCJPN_l01_n02_HRB0071
        0.5%
        42%
        0.6
        000000000-JCJPN_l01_n02_HRB0084
        1.5%
        45%
        0.6
        000000000-JCJPN_l01_n02_HRB0092
        0.1%
        50%
        0.1
        000000000-JCJPN_l01_n02_HRB0093
        1.0%
        42%
        0.6
        000000000-JCJPN_l01_n02_HRB0094
        1.1%
        42%
        0.8
        000000000-JCJPN_l01_n02_HRB0171
        1.4%
        45%
        0.6
        000000000-JCJPN_l01_n02_HRB0172
        1.2%
        46%
        0.9
        000000000-JCJPN_l01_n02_HRB0192
        0.7%
        41%
        0.5
        000000000-JCJPN_l01_n02_HRB0203
        0.7%
        45%
        0.6
        000000000-JCJPN_l01_n02_HRB0204
        0.6%
        45%
        0.7
        000000000-JCJPN_l01_n02_HRB0206
        2.5%
        44%
        0.1
        000000000-JCJPN_l01_n02_HRB0208
        0.7%
        44%
        0.5
        000000000-JCJPN_l01_n02_HRB0209
        0.6%
        43%
        0.7
        000000000-JCJPN_l01_n02_HRB0210
        0.3%
        43%
        0.4
        000000000-JCJPN_l01_n02_HRB0211
        0.6%
        42%
        0.4
        000000000-JCJPN_l01_n02_HRB0249
        1.1%
        46%
        0.7
        000000000-JCJPN_l01_n02_HRB0250
        1.8%
        43%
        0.8
        000000000-JCJPN_l01_n02_HRB0256
        0.5%
        42%
        0.6
        000000000-JCJPN_l01_n02_HRB0257
        0.8%
        41%
        0.7
        000000000-JCJPN_l01_n02_HRB0258L
        0.2%
        44%
        0.3
        000000000-JCJPN_l01_n02_HRB0258R
        0.2%
        46%
        0.3
        000000000-JCJPN_l01_n02_HRB0259L
        0.5%
        42%
        0.3
        000000000-JCJPN_l01_n02_HRB0259R
        0.1%
        51%
        0.3
        000000000-JCJPN_l01_n02_HRB0260L
        0.2%
        43%
        0.3
        000000000-JCJPN_l01_n02_HRB0260R
        0.1%
        49%
        0.4
        000000000-JCJPN_l01_n02_HRB0261L
        0.4%
        42%
        0.3
        000000000-JCJPN_l01_n02_HRB0261R
        0.2%
        45%
        0.4
        000000000-JCJPN_l01_n02_HRB0263L
        0.4%
        43%
        0.5
        000000000-JCJPN_l01_n02_HRB0263R
        0.1%
        48%
        0.5
        000000000-JCJPN_l01_n02_HRB0268
        0.4%
        42%
        0.3
        000000000-JCJPN_l01_n02_HRB0269
        4.0%
        39%
        0.4
        000000000-JCJPN_l01_n02_HRB0270
        1.2%
        41%
        0.3
        000000000-JCJPN_l01_n02_HRB0271
        0.3%
        41%
        0.3
        000000000-JCJPN_l01_n02_HRB0272
        0.5%
        42%
        0.7
        000000000-JCJPN_l01_n02_HRB0273
        0.5%
        42%
        0.1
        000000000-JCJPN_l01_n02_HRBB1EB
        0.1%
        46%
        0.0
        000000000-JCJPN_l01_n02_HRBB1LB
        0.0%
        44%
        0.0
        000000000-JCJPN_l01_n02_HRBB2EB
        0.9%
        45%
        0.0
        000000000-JCJPN_l01_n02_HRBB2LB
        7.0%
        43%
        0.0
        000000000-JCJPN_l01_n02_HRBB3EB
        2.6%
        48%
        0.0
        000000000-JCJPN_l01_n02_HRBB3LB
        4.4%
        44%
        0.0
        000000000-JCJPN_l01_n02_HRBB4EB
        0.2%
        47%
        0.0
        000000000-JCJPN_l01_n02_HRBB4LB
        13.0%
        42%
        0.0
        000000000-JCJPN_l01_n02_undetermined
        58.6%
        44%
        0.7

        Lane Statistics

        Showing 1/1 rows and 4/4 columns.
        LaneTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        1.0
        21555020
        19508841
        3.3
        2.1

        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 48/48 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        650443
        3.3
        HRB0171
        608447
        3.1
        HRB0172
        888570
        4.6
        HRB0203
        596882
        3.1
        HRB0249
        651533
        3.3
        HRB0258L
        266711
        1.4
        HRB0259L
        305957
        1.6
        HRB0268
        267459
        1.4
        HRB0270
        345063
        1.8
        HRB0258R
        257467
        1.3
        HRB0259R
        270305
        1.4
        HRBB1EB
        777.0
        0.0
        HRBB1LB
        125.0
        0.0
        HRB0192
        464540
        2.4
        HRB0204
        652202
        3.3
        HRB0206
        71873
        0.4
        HRB0250
        805740
        4.1
        HRB0260L
        299262
        1.5
        HRB0260R
        362640
        1.9
        HRB0261L
        338982
        1.7
        HRB0261R
        397633
        2.0
        HRB0269
        429031
        2.2
        HRB0271
        344979
        1.8
        HRBB2EB
        11333
        0.1
        HRBB2LB
        1443
        0.0
        HRB0208
        491886
        2.5
        HRB0209
        717807
        3.7
        HRB0210
        409412
        2.1
        HRB0211
        445394
        2.3
        HRB0256
        560592
        2.9
        HRB0257
        685750
        3.5
        HRB0263L
        519691
        2.7
        HRB0272
        741868
        3.8
        HRB0273
        94075
        0.5
        HRB0263R
        535234
        2.7
        HRBB3EB
        1999
        0.0
        HRBB3LB
        435.0
        0.0
        HRB0060
        553630
        2.8
        HRB0062
        655884
        3.4
        HRB0069
        587897
        3.0
        HRB0070
        497230
        2.5
        HRB0071
        646912
        3.3
        HRB0084
        580075
        3.0
        HRB0092
        52050
        0.3
        HRB0093
        594590
        3.0
        HRB0094
        846574
        4.3
        HRBB4EB
        436.0
        0.0
        HRBB4LB
        23.0
        0.0

        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 (%)
        TACTCGCTCTTTCC
        9622.0
        1.5
        CGATGTATCTTTCC
        6813.0
        1.0
        ATCCTCTTCTTTCC
        6471.0
        1.0
        CCTTAATTCTTTCC
        4932.0
        0.8
        CTATCTTTCTTTCC
        4895.0
        0.8
        CTCCTCTTCTTTCC
        4727.0
        0.7
        CTCCTCCTCTTTCC
        4156.0
        0.6
        AGCGCCATCTTTCC
        4049.0
        0.6
        CGCCAACTCTTTCC
        3926.0
        0.6
        CCTTCCTTCTTTCC
        3694.0
        0.6
        CTCTCTTTCTTTCC
        3587.0
        0.6
        CCTAACGTCTTTCC
        3260.0
        0.5
        CTCCCCCTCTTTCC
        3088.0
        0.5
        TCTCCTATCTTTCC
        2918.0
        0.5
        CCTCCCTTCTTTCC
        2863.0
        0.4
        TCCTCTCTCTTTCC
        2832.0
        0.4
        ATCATAATCTTTCC
        2778.0
        0.4
        AGACTCCTCTTTCC
        2760.0
        0.4
        TACTCTCTCTTTCC
        2241.0
        0.3
        TCTCCTCTCTTTCC
        2207.0
        0.3

        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 (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.

        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..