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        Note that additional data was saved in multiqc_data when this report was generated.


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        If you use plots from MultiQC in a publication or presentation, please cite:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

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        About MultiQC

        This report was generated using MultiQC, version 1.9

        You can see a YouTube video describing how to use MultiQC reports here: https://youtu.be/qPbIlO_KWN0

        For more information about MultiQC, including other videos and extensive documentation, please visit http://multiqc.info

        You can report bugs, suggest improvements and find the source code for MultiQC on GitHub: https://github.com/ewels/MultiQC

        MultiQC is published in Bioinformatics:

        MultiQC: Summarize analysis results for multiple tools and samples in a single report
        Philip Ewels, Måns Magnusson, Sverker Lundin and Max Käller
        Bioinformatics (2016)
        doi: 10.1093/bioinformatics/btw354
        PMID: 27312411

        A modular tool to aggregate results from bioinformatics analyses across many samples into a single report.

        Report generated on 2022-04-08, 11:25 based on data in: /scratch/gencore/logs/html/HVCF2DRXY/1


        General Statistics

        Showing 97/97 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HVCF2DRXY_l01_n01_WW100
        23.0%
        38%
        8.0
        HVCF2DRXY_l01_n01_WW102
        22.7%
        38%
        9.5
        HVCF2DRXY_l01_n01_WW103
        22.6%
        37%
        8.7
        HVCF2DRXY_l01_n01_WW107
        21.9%
        38%
        6.4
        HVCF2DRXY_l01_n01_WW108
        23.1%
        38%
        9.2
        HVCF2DRXY_l01_n01_WW109
        24.3%
        37%
        9.4
        HVCF2DRXY_l01_n01_WW111
        23.7%
        38%
        8.4
        HVCF2DRXY_l01_n01_WW112
        21.4%
        38%
        7.1
        HVCF2DRXY_l01_n01_WW113
        24.1%
        38%
        8.8
        HVCF2DRXY_l01_n01_WW114
        19.6%
        38%
        3.8
        HVCF2DRXY_l01_n01_WW115
        19.2%
        38%
        4.0
        HVCF2DRXY_l01_n01_WW116
        22.2%
        38%
        5.2
        HVCF2DRXY_l01_n01_WW117
        21.4%
        36%
        5.6
        HVCF2DRXY_l01_n01_WW118
        22.6%
        37%
        7.0
        HVCF2DRXY_l01_n01_WW119
        20.7%
        37%
        5.1
        HVCF2DRXY_l01_n01_WW120
        24.1%
        37%
        8.2
        HVCF2DRXY_l01_n01_WW121
        23.3%
        37%
        8.4
        HVCF2DRXY_l01_n01_WW122
        23.7%
        37%
        8.2
        HVCF2DRXY_l01_n01_WW123
        20.7%
        37%
        4.9
        HVCF2DRXY_l01_n01_WW124
        22.0%
        37%
        6.8
        HVCF2DRXY_l01_n01_WW125
        23.6%
        37%
        12.3
        HVCF2DRXY_l01_n01_WW126
        21.8%
        37%
        7.3
        HVCF2DRXY_l01_n01_WW127
        22.7%
        37%
        8.0
        HVCF2DRXY_l01_n01_WW128
        20.5%
        37%
        5.4
        HVCF2DRXY_l01_n01_WW129
        22.9%
        37%
        6.5
        HVCF2DRXY_l01_n01_WW130
        21.7%
        37%
        6.7
        HVCF2DRXY_l01_n01_WW132
        22.5%
        37%
        9.0
        HVCF2DRXY_l01_n01_WW133
        25.0%
        36%
        10.0
        HVCF2DRXY_l01_n01_WW134
        24.1%
        38%
        5.7
        HVCF2DRXY_l01_n01_WW135
        26.1%
        37%
        11.0
        HVCF2DRXY_l01_n01_WW137
        23.3%
        36%
        9.0
        HVCF2DRXY_l01_n01_WW138
        23.8%
        37%
        8.0
        HVCF2DRXY_l01_n01_WW139
        21.2%
        37%
        9.8
        HVCF2DRXY_l01_n01_WW143
        21.3%
        38%
        7.9
        HVCF2DRXY_l01_n01_WW144
        22.3%
        37%
        8.9
        HVCF2DRXY_l01_n01_WW145
        22.4%
        37%
        9.3
        HVCF2DRXY_l01_n01_WW146
        23.6%
        37%
        7.5
        HVCF2DRXY_l01_n01_WW147
        23.1%
        37%
        8.1
        HVCF2DRXY_l01_n01_WW148
        23.7%
        37%
        8.3
        HVCF2DRXY_l01_n01_WW149
        21.8%
        37%
        8.4
        HVCF2DRXY_l01_n01_WW150
        21.3%
        36%
        7.0
        HVCF2DRXY_l01_n01_WW151
        22.5%
        37%
        7.8
        HVCF2DRXY_l01_n01_WW152
        21.8%
        37%
        8.1
        HVCF2DRXY_l01_n01_WW153
        25.4%
        36%
        10.5
        HVCF2DRXY_l01_n01_WW154
        20.4%
        36%
        9.0
        HVCF2DRXY_l01_n01_WW155
        21.7%
        36%
        8.5
        HVCF2DRXY_l01_n01_WW156
        22.8%
        37%
        7.6
        HVCF2DRXY_l01_n01_WW157
        23.2%
        36%
        9.6
        HVCF2DRXY_l01_n01_WW158
        22.3%
        37%
        7.7
        HVCF2DRXY_l01_n01_WW159
        25.6%
        36%
        7.7
        HVCF2DRXY_l01_n01_WW160
        24.9%
        36%
        8.3
        HVCF2DRXY_l01_n01_WW161
        22.5%
        36%
        9.0
        HVCF2DRXY_l01_n01_WW162
        22.1%
        36%
        6.6
        HVCF2DRXY_l01_n01_WW163
        19.6%
        36%
        4.7
        HVCF2DRXY_l01_n01_WW164
        18.5%
        36%
        5.2
        HVCF2DRXY_l01_n01_WW165
        21.4%
        36%
        6.9
        HVCF2DRXY_l01_n01_WW167
        21.2%
        37%
        9.4
        HVCF2DRXY_l01_n01_WW168
        21.7%
        36%
        9.3
        HVCF2DRXY_l01_n01_WW169
        21.6%
        36%
        9.3
        HVCF2DRXY_l01_n01_WW170
        26.3%
        37%
        27.0
        HVCF2DRXY_l01_n01_WW51
        21.4%
        37%
        5.5
        HVCF2DRXY_l01_n01_WW52
        21.0%
        39%
        5.1
        HVCF2DRXY_l01_n01_WW53
        23.6%
        37%
        7.7
        HVCF2DRXY_l01_n01_WW54
        20.0%
        37%
        4.5
        HVCF2DRXY_l01_n01_WW55
        22.4%
        37%
        8.7
        HVCF2DRXY_l01_n01_WW56
        19.0%
        38%
        4.4
        HVCF2DRXY_l01_n01_WW57
        19.1%
        37%
        4.7
        HVCF2DRXY_l01_n01_WW58
        23.1%
        37%
        6.7
        HVCF2DRXY_l01_n01_WW59
        21.8%
        36%
        7.0
        HVCF2DRXY_l01_n01_WW62
        22.2%
        38%
        7.9
        HVCF2DRXY_l01_n01_WW65
        23.7%
        38%
        9.0
        HVCF2DRXY_l01_n01_WW66
        24.0%
        38%
        13.6
        HVCF2DRXY_l01_n01_WW67
        21.7%
        38%
        6.8
        HVCF2DRXY_l01_n01_WW68
        24.4%
        37%
        8.1
        HVCF2DRXY_l01_n01_WW70
        22.5%
        38%
        6.0
        HVCF2DRXY_l01_n01_WW71
        22.4%
        38%
        7.4
        HVCF2DRXY_l01_n01_WW72
        22.6%
        37%
        7.6
        HVCF2DRXY_l01_n01_WW74
        24.3%
        37%
        9.2
        HVCF2DRXY_l01_n01_WW75
        23.7%
        37%
        12.0
        HVCF2DRXY_l01_n01_WW76
        21.4%
        37%
        8.3
        HVCF2DRXY_l01_n01_WW77
        24.0%
        37%
        11.5
        HVCF2DRXY_l01_n01_WW78
        23.2%
        37%
        7.1
        HVCF2DRXY_l01_n01_WW80
        22.9%
        36%
        6.7
        HVCF2DRXY_l01_n01_WW81
        23.3%
        37%
        7.6
        HVCF2DRXY_l01_n01_WW82
        23.5%
        37%
        7.5
        HVCF2DRXY_l01_n01_WW83
        24.5%
        37%
        9.9
        HVCF2DRXY_l01_n01_WW87
        24.4%
        38%
        7.8
        HVCF2DRXY_l01_n01_WW88
        22.1%
        37%
        7.0
        HVCF2DRXY_l01_n01_WW89
        22.6%
        37%
        7.6
        HVCF2DRXY_l01_n01_WW90
        23.4%
        37%
        8.4
        HVCF2DRXY_l01_n01_WW91
        21.4%
        37%
        6.5
        HVCF2DRXY_l01_n01_WW93
        25.0%
        38%
        8.4
        HVCF2DRXY_l01_n01_WW95
        24.3%
        37%
        9.4
        HVCF2DRXY_l01_n01_WW96
        24.8%
        38%
        8.8
        HVCF2DRXY_l01_n01_WW97
        26.5%
        37%
        10.3
        HVCF2DRXY_l01_n01_WW99
        24.2%
        38%
        8.2
        HVCF2DRXY_l01_n01_undetermined
        94.0%
        45%
        219.5

        Lane Statistics

        Showing 1/1 rows and 4/4 columns.
        LaneTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        1.0
        1276674048
        991239569
        22.1
        20.2

        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 97/97 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        219522777
        22.1
        WW51
        5451252
        0.6
        WW52
        5137053
        0.5
        WW53
        7737052
        0.8
        WW54
        4513121
        0.5
        WW55
        8690728
        0.9
        WW56
        4372840
        0.4
        WW57
        4676219
        0.5
        WW58
        6736398
        0.7
        WW59
        6991380
        0.7
        WW62
        7857029
        0.8
        WW65
        9015473
        0.9
        WW66
        13578687
        1.4
        WW67
        6819680
        0.7
        WW68
        8108035
        0.8
        WW70
        5967268
        0.6
        WW71
        7353617
        0.7
        WW72
        7629802
        0.8
        WW74
        9186070
        0.9
        WW75
        11963795
        1.2
        WW76
        8320536
        0.8
        WW77
        11465848
        1.2
        WW78
        7087478
        0.7
        WW80
        6738797
        0.7
        WW81
        7567899
        0.8
        WW82
        7500544
        0.8
        WW83
        9866657
        1.0
        WW87
        7754605
        0.8
        WW88
        6967651
        0.7
        WW89
        7553307
        0.8
        WW90
        8361561
        0.8
        WW91
        6524194
        0.7
        WW93
        8438659
        0.9
        WW95
        9399112
        0.9
        WW96
        8845185
        0.9
        WW97
        10333719
        1.0
        WW99
        8217336
        0.8
        WW100
        7969085
        0.8
        WW102
        9480734
        1.0
        WW103
        8670046
        0.9
        WW107
        6385089
        0.6
        WW108
        9173770
        0.9
        WW109
        9417695
        0.9
        WW111
        8373713
        0.8
        WW112
        7127809
        0.7
        WW113
        8842922
        0.9
        WW114
        3789047
        0.4
        WW115
        3973591
        0.4
        WW116
        5230812
        0.5
        WW117
        5559505
        0.6
        WW118
        6951720
        0.7
        WW119
        5110850
        0.5
        WW120
        8182469
        0.8
        WW121
        8377774
        0.8
        WW122
        8248876
        0.8
        WW123
        4922562
        0.5
        WW124
        6836265
        0.7
        WW125
        12284393
        1.2
        WW126
        7261158
        0.7
        WW127
        8018112
        0.8
        WW128
        5388899
        0.5
        WW129
        6503272
        0.7
        WW130
        6730497
        0.7
        WW132
        9041192
        0.9
        WW133
        10002854
        1.0
        WW134
        5716412
        0.6
        WW135
        10957581
        1.1
        WW137
        8973920
        0.9
        WW138
        7963858
        0.8
        WW139
        9813934
        1.0
        WW143
        7855669
        0.8
        WW144
        8873882
        0.9
        WW145
        9288728
        0.9
        WW146
        7470543
        0.8
        WW147
        8053015
        0.8
        WW148
        8277766
        0.8
        WW149
        8398381
        0.8
        WW150
        6985382
        0.7
        WW151
        7826200
        0.8
        WW152
        8129319
        0.8
        WW153
        10510734
        1.1
        WW154
        9013119
        0.9
        WW155
        8530979
        0.9
        WW156
        7643501
        0.8
        WW157
        9638993
        1.0
        WW158
        7729491
        0.8
        WW159
        7697402
        0.8
        WW160
        8298126
        0.8
        WW161
        9002145
        0.9
        WW162
        6597242
        0.7
        WW163
        4666061
        0.5
        WW164
        5227545
        0.5
        WW165
        6931724
        0.7
        WW167
        9447665
        1.0
        WW168
        9263335
        0.9
        WW169
        9337839
        0.9
        WW170
        27013003
        2.7

        Barcodes of Undetermined Reads


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

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        GGGGGG
        213000180.0
        97.0
        AAAAAA
        150428.0
        0.1
        CGGGGG
        85321.0
        0.0
        AAACAA
        81316.0
        0.0
        AAAAAC
        75113.0
        0.0
        ACAAAA
        72237.0
        0.0
        CAAAAA
        68405.0
        0.0
        GGGGGC
        61447.0
        0.0
        AAAATA
        55341.0
        0.0
        AAATAA
        52752.0
        0.0
        AAAAAT
        51122.0
        0.0
        GGGGTG
        49775.0
        0.0
        AGAAAA
        46739.0
        0.0
        GGGGCG
        44807.0
        0.0
        ACAAAC
        40803.0
        0.0
        ACACAA
        40494.0
        0.0
        AAAACC
        36726.0
        0.0
        TAACTA
        35925.0
        0.0
        TGGTAA
        34071.0
        0.0
        AAACAC
        32944.0
        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.

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        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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