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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 2022-03-23, 06:31 based on data in: /scratch/gencore/logs/html/HMYYNDRXY/2


        General Statistics

        Showing 97/97 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        HMYYNDRXY_l02_n01_RS147
        18.0%
        37%
        7.1
        HMYYNDRXY_l02_n01_RS148
        25.6%
        38%
        8.4
        HMYYNDRXY_l02_n01_RS149
        20.2%
        37%
        8.9
        HMYYNDRXY_l02_n01_RS150
        19.7%
        37%
        8.9
        HMYYNDRXY_l02_n01_RS151
        22.9%
        37%
        12.7
        HMYYNDRXY_l02_n01_RS152
        19.5%
        37%
        8.2
        HMYYNDRXY_l02_n01_RS153
        18.6%
        37%
        8.5
        HMYYNDRXY_l02_n01_RS154
        25.1%
        37%
        9.9
        HMYYNDRXY_l02_n01_RS155
        23.6%
        38%
        9.3
        HMYYNDRXY_l02_n01_RS156
        22.4%
        37%
        8.5
        HMYYNDRXY_l02_n01_RS157
        24.0%
        38%
        8.0
        HMYYNDRXY_l02_n01_RS158
        22.1%
        38%
        9.8
        HMYYNDRXY_l02_n01_RS159
        19.5%
        37%
        8.7
        HMYYNDRXY_l02_n01_RS160
        21.3%
        38%
        8.7
        HMYYNDRXY_l02_n01_RS161
        18.5%
        37%
        9.1
        HMYYNDRXY_l02_n01_RS162
        19.7%
        37%
        8.7
        HMYYNDRXY_l02_n01_RS163
        20.8%
        36%
        8.8
        HMYYNDRXY_l02_n01_RS164
        21.2%
        35%
        8.9
        HMYYNDRXY_l02_n01_RS165
        20.5%
        37%
        9.4
        HMYYNDRXY_l02_n01_RS166
        19.5%
        37%
        8.3
        HMYYNDRXY_l02_n01_RS167
        21.5%
        37%
        10.9
        HMYYNDRXY_l02_n01_RS168
        25.8%
        37%
        9.2
        HMYYNDRXY_l02_n01_RS169
        21.0%
        37%
        9.4
        HMYYNDRXY_l02_n01_RS170
        18.5%
        37%
        8.9
        HMYYNDRXY_l02_n01_RS171
        19.8%
        37%
        9.4
        HMYYNDRXY_l02_n01_RS172
        20.1%
        37%
        6.6
        HMYYNDRXY_l02_n01_RS173
        19.7%
        37%
        9.0
        HMYYNDRXY_l02_n01_RS174
        22.3%
        37%
        8.0
        HMYYNDRXY_l02_n01_RS175
        19.5%
        36%
        9.9
        HMYYNDRXY_l02_n01_RS176
        24.0%
        37%
        9.8
        HMYYNDRXY_l02_n01_RS177
        20.6%
        37%
        9.5
        HMYYNDRXY_l02_n01_RS178
        18.7%
        37%
        8.5
        HMYYNDRXY_l02_n01_RS179
        21.1%
        36%
        8.2
        HMYYNDRXY_l02_n01_RS180
        18.8%
        37%
        9.2
        HMYYNDRXY_l02_n01_RS181
        20.2%
        37%
        9.9
        HMYYNDRXY_l02_n01_RS182
        18.8%
        37%
        8.1
        HMYYNDRXY_l02_n01_RS183
        22.8%
        37%
        9.8
        HMYYNDRXY_l02_n01_RS184
        17.9%
        37%
        8.7
        HMYYNDRXY_l02_n01_RS185
        18.7%
        37%
        8.4
        HMYYNDRXY_l02_n01_RS186
        18.8%
        36%
        8.6
        HMYYNDRXY_l02_n01_RS187
        19.2%
        36%
        9.2
        HMYYNDRXY_l02_n01_RS188
        19.5%
        36%
        11.4
        HMYYNDRXY_l02_n01_RS189
        19.2%
        37%
        9.6
        HMYYNDRXY_l02_n01_RS190
        21.0%
        36%
        11.4
        HMYYNDRXY_l02_n01_RS191
        20.0%
        37%
        10.4
        HMYYNDRXY_l02_n01_RS192
        18.6%
        36%
        8.3
        HMYYNDRXY_l02_n01_RS193
        19.2%
        37%
        9.8
        HMYYNDRXY_l02_n01_RS194
        24.1%
        36%
        10.2
        HMYYNDRXY_l02_n01_RS195
        21.5%
        37%
        9.1
        HMYYNDRXY_l02_n01_RS196
        18.8%
        37%
        7.7
        HMYYNDRXY_l02_n01_RS197
        20.1%
        37%
        9.6
        HMYYNDRXY_l02_n01_RS198
        21.6%
        37%
        10.6
        HMYYNDRXY_l02_n01_RS199
        22.2%
        37%
        11.7
        HMYYNDRXY_l02_n01_RS200
        21.0%
        38%
        9.9
        HMYYNDRXY_l02_n01_RS201
        22.0%
        38%
        10.7
        HMYYNDRXY_l02_n01_RS202
        23.9%
        38%
        10.7
        HMYYNDRXY_l02_n01_RS203
        20.6%
        37%
        9.9
        HMYYNDRXY_l02_n01_RS205
        20.0%
        38%
        8.9
        HMYYNDRXY_l02_n01_RS206
        22.2%
        37%
        9.5
        HMYYNDRXY_l02_n01_RS213
        20.7%
        37%
        9.0
        HMYYNDRXY_l02_n01_RS260
        17.9%
        37%
        9.2
        HMYYNDRXY_l02_n01_WW1
        20.0%
        38%
        6.3
        HMYYNDRXY_l02_n01_WW14
        22.9%
        38%
        10.5
        HMYYNDRXY_l02_n01_WW15
        21.8%
        38%
        8.3
        HMYYNDRXY_l02_n01_WW16
        19.8%
        38%
        8.4
        HMYYNDRXY_l02_n01_WW17
        22.0%
        37%
        9.0
        HMYYNDRXY_l02_n01_WW18
        21.3%
        38%
        8.6
        HMYYNDRXY_l02_n01_WW19
        22.1%
        37%
        10.6
        HMYYNDRXY_l02_n01_WW2
        21.6%
        38%
        9.7
        HMYYNDRXY_l02_n01_WW20
        24.1%
        37%
        9.7
        HMYYNDRXY_l02_n01_WW21
        21.2%
        37%
        10.3
        HMYYNDRXY_l02_n01_WW22
        22.7%
        37%
        9.5
        HMYYNDRXY_l02_n01_WW23
        19.1%
        37%
        7.8
        HMYYNDRXY_l02_n01_WW24
        22.9%
        37%
        9.0
        HMYYNDRXY_l02_n01_WW27
        22.5%
        37%
        11.3
        HMYYNDRXY_l02_n01_WW28
        21.3%
        37%
        10.0
        HMYYNDRXY_l02_n01_WW3
        24.2%
        37%
        13.1
        HMYYNDRXY_l02_n01_WW31
        20.7%
        38%
        9.1
        HMYYNDRXY_l02_n01_WW35
        21.5%
        37%
        9.1
        HMYYNDRXY_l02_n01_WW37
        18.5%
        38%
        7.8
        HMYYNDRXY_l02_n01_WW38
        23.0%
        38%
        10.7
        HMYYNDRXY_l02_n01_WW41
        21.3%
        37%
        9.8
        HMYYNDRXY_l02_n01_WW42
        19.8%
        37%
        9.0
        HMYYNDRXY_l02_n01_WW43
        22.9%
        37%
        11.9
        HMYYNDRXY_l02_n01_WW44
        21.7%
        37%
        8.6
        HMYYNDRXY_l02_n01_WW45
        21.0%
        37%
        8.7
        HMYYNDRXY_l02_n01_WW46
        20.5%
        37%
        9.9
        HMYYNDRXY_l02_n01_WW47
        20.0%
        37%
        9.1
        HMYYNDRXY_l02_n01_WW48
        21.6%
        36%
        8.9
        HMYYNDRXY_l02_n01_WW49
        20.9%
        38%
        10.1
        HMYYNDRXY_l02_n01_WW5
        21.2%
        37%
        9.5
        HMYYNDRXY_l02_n01_WW50
        20.6%
        37%
        8.8
        HMYYNDRXY_l02_n01_WW6
        26.7%
        37%
        20.7
        HMYYNDRXY_l02_n01_WW7
        21.3%
        38%
        9.0
        HMYYNDRXY_l02_n01_WW8
        21.5%
        38%
        9.5
        HMYYNDRXY_l02_n01_WW9
        20.7%
        37%
        11.0
        HMYYNDRXY_l02_n01_undetermined
        94.3%
        46%
        196.1

        Lane Statistics

        Showing 1/1 rows and 4/4 columns.
        LaneTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        2.0
        1276674048
        1104597014
        17.8
        16.1

        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 97/97 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        196110212
        17.8
        RS147
        7077364
        0.6
        RS148
        8371959
        0.8
        RS149
        8933432
        0.8
        RS150
        8883110
        0.8
        RS151
        12706147
        1.1
        RS152
        8166431
        0.7
        RS153
        8452359
        0.8
        RS154
        9906311
        0.9
        RS155
        9287436
        0.8
        RS156
        8467831
        0.8
        RS157
        7977745
        0.7
        RS158
        9816808
        0.9
        RS159
        8673965
        0.8
        RS160
        8660144
        0.8
        RS161
        9062796
        0.8
        RS162
        8658681
        0.8
        RS163
        8750650
        0.8
        RS164
        8905762
        0.8
        RS165
        9361345
        0.8
        RS166
        8345007
        0.8
        RS167
        10896636
        1.0
        RS168
        9154062
        0.8
        RS169
        9420026
        0.9
        RS170
        8870750
        0.8
        RS171
        9383140
        0.8
        RS172
        6606172
        0.6
        RS173
        9020803
        0.8
        RS174
        8041925
        0.7
        RS175
        9907411
        0.9
        RS176
        9829435
        0.9
        RS177
        9484215
        0.9
        RS178
        8460452
        0.8
        RS179
        8188167
        0.7
        RS180
        9224925
        0.8
        RS181
        9887703
        0.9
        RS182
        8130779
        0.7
        RS183
        9761744
        0.9
        RS184
        8657595
        0.8
        RS185
        8434039
        0.8
        RS186
        8637527
        0.8
        RS187
        9161320
        0.8
        RS188
        11390008
        1.0
        RS189
        9589481
        0.9
        RS190
        11402524
        1.0
        RS191
        10352483
        0.9
        RS192
        8339270
        0.8
        RS193
        9766107
        0.9
        RS194
        10219046
        0.9
        RS195
        9120188
        0.8
        RS196
        7659825
        0.7
        RS197
        9559570
        0.9
        RS198
        10589287
        1.0
        RS199
        11706040
        1.1
        RS200
        9936639
        0.9
        RS201
        10659941
        1.0
        RS202
        10711852
        1.0
        RS203
        9942235
        0.9
        RS205
        8903488
        0.8
        RS206
        9455518
        0.9
        RS213
        8993212
        0.8
        RS260
        9219381
        0.8
        WW1
        6295375
        0.6
        WW2
        9663219
        0.9
        WW3
        13114620
        1.2
        WW5
        9458389
        0.9
        WW6
        20693060
        1.9
        WW7
        8961491
        0.8
        WW8
        9492665
        0.9
        WW9
        11022371
        1.0
        WW14
        10513065
        1.0
        WW15
        8301180
        0.8
        WW16
        8423903
        0.8
        WW17
        8977722
        0.8
        WW18
        8581273
        0.8
        WW19
        10576728
        1.0
        WW20
        9704065
        0.9
        WW21
        10301746
        0.9
        WW22
        9506608
        0.9
        WW23
        7837854
        0.7
        WW24
        9030510
        0.8
        WW27
        11337653
        1.0
        WW28
        9995591
        0.9
        WW31
        9129422
        0.8
        WW35
        9125064
        0.8
        WW37
        7846245
        0.7
        WW38
        10747671
        1.0
        WW41
        9759318
        0.9
        WW42
        9027343
        0.8
        WW43
        11860409
        1.1
        WW44
        8607014
        0.8
        WW45
        8704920
        0.8
        WW46
        9889700
        0.9
        WW47
        9129783
        0.8
        WW48
        8855399
        0.8
        WW49
        10098695
        0.9
        WW50
        8776527
        0.8

        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
        190821060.0
        97.3
        CGGGGG
        103663.0
        0.1
        AAAAAA
        97924.0
        0.1
        GGGGGC
        56082.0
        0.0
        NGGGGG
        47016.0
        0.0
        AAACAA
        44575.0
        0.0
        CAAAAA
        44333.0
        0.0
        TTATGA
        42537.0
        0.0
        TGATTA
        39111.0
        0.0
        GGGGTG
        38246.0
        0.0
        AAAAAC
        37020.0
        0.0
        ACAAAA
        36254.0
        0.0
        ACCGAA
        34612.0
        0.0
        GGGGCG
        33938.0
        0.0
        TGGTAA
        33879.0
        0.0
        AAATAA
        31939.0
        0.0
        AAAATA
        31596.0
        0.0
        ATTGTA
        31368.0
        0.0
        CTGCTA
        30931.0
        0.0
        AAGCAA
        30262.0
        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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