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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 2022-04-08, 12:20 based on data in: /scratch/gencore/logs/html/HVCF2DRXY/2


        General Statistics

        Showing 78/78 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HVCF2DRXY_l02_n01_2hR1_KCl
        20.7%
        37%
        9.5
        HVCF2DRXY_l02_n01_2hR1_N
        20.9%
        38%
        11.6
        HVCF2DRXY_l02_n01_2hR2_KCl
        21.3%
        38%
        12.2
        HVCF2DRXY_l02_n01_2hR2_N
        21.0%
        39%
        11.1
        HVCF2DRXY_l02_n01_2hR3_KCl
        21.1%
        38%
        13.5
        HVCF2DRXY_l02_n01_2hR3_N
        21.8%
        39%
        13.5
        HVCF2DRXY_l02_n01_2hS1_KCl
        23.4%
        38%
        12.8
        HVCF2DRXY_l02_n01_2hS1_N
        22.5%
        38%
        12.1
        HVCF2DRXY_l02_n01_2hS2_KCl
        23.1%
        39%
        13.7
        HVCF2DRXY_l02_n01_2hS2_N
        22.3%
        38%
        12.0
        HVCF2DRXY_l02_n01_2hS3_KCl
        24.4%
        38%
        13.9
        HVCF2DRXY_l02_n01_2hS3_N
        21.8%
        38%
        10.5
        HVCF2DRXY_l02_n01_RS110_re
        43.7%
        39%
        8.2
        HVCF2DRXY_l02_n01_RS32_re
        32.6%
        37%
        14.9
        HVCF2DRXY_l02_n01_WW171
        24.9%
        37%
        10.8
        HVCF2DRXY_l02_n01_WW172
        26.7%
        38%
        12.2
        HVCF2DRXY_l02_n01_WW173
        24.7%
        37%
        10.5
        HVCF2DRXY_l02_n01_WW174
        23.3%
        37%
        6.4
        HVCF2DRXY_l02_n01_WW175
        21.9%
        37%
        6.1
        HVCF2DRXY_l02_n01_WW176
        25.7%
        37%
        10.4
        HVCF2DRXY_l02_n01_WW177
        21.1%
        37%
        5.1
        HVCF2DRXY_l02_n01_WW178
        23.4%
        38%
        5.6
        HVCF2DRXY_l02_n01_WW179
        20.8%
        38%
        4.0
        HVCF2DRXY_l02_n01_WW180
        19.4%
        38%
        4.8
        HVCF2DRXY_l02_n01_WW181
        27.0%
        38%
        14.9
        HVCF2DRXY_l02_n01_WW182
        22.6%
        38%
        6.0
        HVCF2DRXY_l02_n01_WW183
        22.2%
        38%
        6.9
        HVCF2DRXY_l02_n01_WW184
        20.8%
        37%
        3.2
        HVCF2DRXY_l02_n01_WW185
        23.7%
        37%
        9.3
        HVCF2DRXY_l02_n01_WW186
        25.3%
        37%
        11.9
        HVCF2DRXY_l02_n01_WW187
        20.9%
        37%
        3.4
        HVCF2DRXY_l02_n01_WW188
        20.6%
        37%
        4.8
        HVCF2DRXY_l02_n01_WW189
        25.4%
        38%
        12.4
        HVCF2DRXY_l02_n01_WW190
        24.1%
        37%
        10.9
        HVCF2DRXY_l02_n01_WW191
        27.0%
        39%
        11.4
        HVCF2DRXY_l02_n01_WW192
        25.6%
        38%
        9.1
        HVCF2DRXY_l02_n01_WW193
        23.5%
        38%
        6.0
        HVCF2DRXY_l02_n01_WW194
        26.6%
        38%
        8.4
        HVCF2DRXY_l02_n01_WW195
        26.9%
        38%
        11.8
        HVCF2DRXY_l02_n01_WW196
        26.6%
        37%
        12.1
        HVCF2DRXY_l02_n01_WW197
        24.3%
        37%
        10.9
        HVCF2DRXY_l02_n01_WW198
        25.0%
        38%
        11.1
        HVCF2DRXY_l02_n01_WW199
        25.6%
        38%
        10.5
        HVCF2DRXY_l02_n01_WW200
        26.5%
        37%
        13.4
        HVCF2DRXY_l02_n01_WW202
        26.9%
        37%
        14.2
        HVCF2DRXY_l02_n01_WW203
        25.3%
        38%
        9.6
        HVCF2DRXY_l02_n01_WW204
        26.1%
        38%
        13.7
        HVCF2DRXY_l02_n01_WW205
        24.5%
        39%
        11.4
        HVCF2DRXY_l02_n01_WW206
        23.5%
        38%
        4.4
        HVCF2DRXY_l02_n01_WW210
        28.5%
        38%
        13.1
        HVCF2DRXY_l02_n01_WW220
        25.4%
        37%
        12.3
        HVCF2DRXY_l02_n01_WW221
        22.8%
        37%
        7.3
        HVCF2DRXY_l02_n01_WW222
        25.8%
        37%
        10.9
        HVCF2DRXY_l02_n01_WW231
        26.4%
        38%
        12.6
        HVCF2DRXY_l02_n01_WW232
        23.0%
        37%
        5.9
        HVCF2DRXY_l02_n01_WW242
        23.4%
        37%
        7.5
        HVCF2DRXY_l02_n01_WW246
        26.6%
        37%
        9.3
        HVCF2DRXY_l02_n01_WW249
        28.5%
        37%
        12.6
        HVCF2DRXY_l02_n01_WW254
        26.1%
        38%
        12.3
        HVCF2DRXY_l02_n01_WW256
        24.2%
        38%
        11.5
        HVCF2DRXY_l02_n01_WW261
        26.5%
        38%
        14.9
        HVCF2DRXY_l02_n01_WW263
        28.9%
        37%
        17.5
        HVCF2DRXY_l02_n01_WW264
        28.0%
        36%
        11.6
        HVCF2DRXY_l02_n01_WW267
        28.4%
        37%
        13.2
        HVCF2DRXY_l02_n01_WW268
        27.1%
        37%
        11.5
        HVCF2DRXY_l02_n01_WW269
        31.0%
        38%
        14.4
        HVCF2DRXY_l02_n01_WW270
        30.9%
        37%
        13.2
        HVCF2DRXY_l02_n01_WW273
        30.8%
        37%
        12.2
        HVCF2DRXY_l02_n01_WW277
        31.0%
        37%
        13.7
        HVCF2DRXY_l02_n01_WW282
        28.0%
        37%
        10.0
        HVCF2DRXY_l02_n01_WW284
        29.0%
        37%
        14.0
        HVCF2DRXY_l02_n01_WW287
        30.9%
        37%
        13.9
        HVCF2DRXY_l02_n01_WW292
        28.7%
        37%
        13.8
        HVCF2DRXY_l02_n01_WW296
        29.6%
        37%
        13.2
        HVCF2DRXY_l02_n01_WW303
        27.7%
        36%
        13.7
        HVCF2DRXY_l02_n01_WW306
        28.6%
        37%
        9.9
        HVCF2DRXY_l02_n01_WW308
        31.7%
        37%
        13.8
        HVCF2DRXY_l02_n01_undetermined
        95.9%
        44%
        280.2

        Lane Statistics

        Showing 1/1 rows and 4/4 columns.
        LaneTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        2.0
        1276674048
        1106960243
        25.3
        24.2

        Lane 2 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 78/78 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        280156266
        25.3
        WW171
        10849538
        1.0
        WW172
        12229661
        1.1
        WW173
        10476908
        0.9
        WW174
        6408246
        0.6
        WW175
        6082108
        0.5
        WW176
        10405830
        0.9
        WW177
        5081297
        0.5
        WW178
        5591329
        0.5
        WW179
        4016825
        0.4
        WW180
        4828421
        0.4
        WW181
        14930316
        1.3
        WW182
        5953125
        0.5
        WW183
        6914091
        0.6
        WW184
        3151379
        0.3
        WW185
        9298178
        0.8
        WW186
        11943750
        1.1
        WW187
        3437219
        0.3
        WW188
        4763610
        0.4
        WW189
        12431883
        1.1
        WW190
        10886807
        1.0
        WW191
        11423672
        1.0
        WW192
        9119562
        0.8
        WW193
        6007547
        0.5
        WW194
        8359664
        0.8
        WW195
        11834490
        1.1
        WW196
        12148692
        1.1
        WW197
        10922609
        1.0
        WW198
        11063145
        1.0
        WW199
        10501707
        0.9
        WW200
        13407203
        1.2
        WW202
        14240848
        1.3
        WW203
        9558365
        0.9
        WW204
        13719259
        1.2
        WW205
        11409772
        1.0
        WW206
        4396322
        0.4
        WW210
        13060640
        1.2
        WW220
        12303724
        1.1
        WW221
        7321915
        0.7
        WW222
        10871246
        1.0
        WW231
        12647205
        1.1
        WW232
        5869373
        0.5
        WW242
        7471740
        0.7
        WW246
        9256003
        0.8
        WW249
        12624690
        1.1
        WW254
        12317610
        1.1
        WW256
        11475406
        1.0
        WW261
        14906892
        1.3
        WW263
        17450230
        1.6
        WW264
        11601985
        1.0
        WW267
        13163927
        1.2
        WW268
        11481361
        1.0
        WW269
        14400323
        1.3
        WW270
        13204496
        1.2
        WW273
        12198212
        1.1
        WW277
        13675260
        1.2
        WW282
        10048114
        0.9
        WW284
        14002610
        1.3
        WW287
        13850154
        1.3
        WW292
        13840138
        1.2
        WW296
        13150251
        1.2
        WW303
        13669206
        1.2
        WW306
        9866405
        0.9
        WW308
        13824159
        1.2
        RS32_re
        14922815
        1.3
        RS110_re
        8199198
        0.7
        2hS1_KCl
        12789965
        1.2
        2hS2_KCl
        13681164
        1.2
        2hS3_KCl
        13893318
        1.3
        2hS1_N
        12051045
        1.1
        2hS2_N
        11965559
        1.1
        2hS3_N
        10540287
        1.0
        2hR1_KCl
        9508983
        0.9
        2hR2_KCl
        12169016
        1.1
        2hR3_KCl
        13539828
        1.2
        2hR1_N
        11624073
        1.1
        2hR2_N
        11106579
        1.0
        2hR3_N
        13465494
        1.2

        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
        273956865.0
        97.8
        CGGGGG
        84626.0
        0.0
        AAAAAA
        68632.0
        0.0
        AACAAG
        55891.0
        0.0
        CAAAAA
        50866.0
        0.0
        GGGGGC
        46416.0
        0.0
        GGGGTG
        46292.0
        0.0
        AACAAA
        44583.0
        0.0
        GGGGCG
        44444.0
        0.0
        TGATTA
        39851.0
        0.0
        TAGTCA
        37421.0
        0.0
        TTATGA
        36663.0
        0.0
        TAACTA
        36601.0
        0.0
        AAAATA
        33688.0
        0.0
        AAATAA
        31582.0
        0.0
        AAGCAA
        29930.0
        0.0
        ATTGTA
        29168.0
        0.0
        ACTAAA
        28707.0
        0.0
        CTGCTA
        28498.0
        0.0
        ACAAAA
        28296.0
        0.0

        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 (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.

        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..