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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 1.7

        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 2019-07-17, 15:17 based on data in: /home/classadmin/class_Summer2019/raw_reads_QC/fastqc_results


        General Statistics

        Showing 112/112 rows and 3/5 columns.
        Sample Name% Dups% GCM Seqs
        SNF2_1 | ERR458500
        32.5%
        42%
        1.9
        SNF2_1 | ERR458501
        31.6%
        42%
        1.9
        SNF2_1 | ERR458502
        32.1%
        42%
        1.9
        SNF2_1 | ERR458503
        32.0%
        42%
        1.7
        SNF2_1 | ERR458504
        30.6%
        42%
        1.5
        SNF2_1 | ERR458505
        29.7%
        42%
        1.5
        SNF2_1 | ERR458506
        34.6%
        42%
        1.9
        SNF2_13 | ERR458584
        29.6%
        41%
        1.3
        SNF2_13 | ERR458585
        29.3%
        41%
        1.3
        SNF2_13 | ERR458586
        28.8%
        41%
        1.3
        SNF2_13 | ERR458587
        28.3%
        41%
        1.2
        SNF2_13 | ERR458588
        28.1%
        41%
        1.0
        SNF2_13 | ERR458589
        27.5%
        41%
        1.0
        SNF2_13 | ERR458590
        31.6%
        41%
        1.3
        SNF2_2 | ERR458507
        33.3%
        43%
        1.6
        SNF2_2 | ERR458508
        32.6%
        43%
        1.6
        SNF2_2 | ERR458509
        32.7%
        43%
        1.6
        SNF2_2 | ERR458510
        32.5%
        43%
        1.4
        SNF2_2 | ERR458511
        31.6%
        43%
        1.2
        SNF2_2 | ERR458512
        30.6%
        43%
        1.2
        SNF2_2 | ERR458513
        35.7%
        43%
        1.6
        SNF2_21 | ERR458640
        33.7%
        42%
        1.8
        SNF2_21 | ERR458641
        32.6%
        42%
        1.8
        SNF2_21 | ERR458642
        33.2%
        42%
        1.7
        SNF2_21 | ERR458643
        33.0%
        42%
        1.6
        SNF2_21 | ERR458644
        31.8%
        42%
        1.4
        SNF2_21 | ERR458645
        31.0%
        42%
        1.4
        SNF2_21 | ERR458646
        35.8%
        42%
        1.8
        SNF2_25 | ERR458668
        37.8%
        44%
        1.6
        SNF2_25 | ERR458669
        37.3%
        44%
        1.5
        SNF2_25 | ERR458670
        37.1%
        44%
        1.5
        SNF2_25 | ERR458671
        36.8%
        44%
        1.4
        SNF2_25 | ERR458672
        36.5%
        44%
        1.2
        SNF2_25 | ERR458673
        35.4%
        44%
        1.2
        SNF2_25 | ERR458674
        40.3%
        44%
        1.6
        SNF2_28 | ERR458689
        34.0%
        43%
        1.5
        SNF2_28 | ERR458690
        33.4%
        43%
        1.5
        SNF2_28 | ERR458691
        33.4%
        43%
        1.5
        SNF2_28 | ERR458692
        33.2%
        43%
        1.3
        SNF2_28 | ERR458693
        32.7%
        43%
        1.2
        SNF2_28 | ERR458694
        31.6%
        43%
        1.2
        SNF2_28 | ERR458695
        36.3%
        43%
        1.5
        SNF2_5 | ERR458528
        33.8%
        43%
        1.4
        SNF2_5 | ERR458529
        33.2%
        43%
        1.4
        SNF2_5 | ERR458530
        32.9%
        43%
        1.4
        SNF2_5 | ERR458531
        32.6%
        43%
        1.2
        SNF2_5 | ERR458532
        32.5%
        43%
        1.1
        SNF2_5 | ERR458533
        31.8%
        43%
        1.1
        SNF2_5 | ERR458534
        35.7%
        43%
        1.4
        SNF2_6 | ERR458535
        37.7%
        42%
        1.6
        SNF2_6 | ERR458536
        37.0%
        41%
        1.6
        SNF2_6 | ERR458537
        36.9%
        41%
        1.6
        SNF2_6 | ERR458538
        36.7%
        41%
        1.4
        SNF2_6 | ERR458539
        36.0%
        41%
        1.2
        SNF2_6 | ERR458540
        34.6%
        41%
        1.2
        SNF2_6 | ERR458541
        40.1%
        41%
        1.6
        WT_1 | ERR458493
        32.3%
        43%
        1.1
        WT_1 | ERR458494
        32.2%
        43%
        1.1
        WT_1 | ERR458495
        31.8%
        43%
        1.1
        WT_1 | ERR458496
        30.9%
        44%
        1.0
        WT_1 | ERR458497
        30.7%
        43%
        0.8
        WT_1 | ERR458498
        30.7%
        43%
        0.9
        WT_1 | ERR458499
        33.9%
        43%
        1.1
        WT_13 | ERR458955
        34.3%
        42%
        1.7
        WT_13 | ERR458956
        33.3%
        42%
        1.7
        WT_13 | ERR458957
        33.5%
        42%
        1.7
        WT_13 | ERR458958
        33.7%
        42%
        1.5
        WT_13 | ERR458959
        32.5%
        42%
        1.3
        WT_13 | ERR458960
        31.6%
        42%
        1.3
        WT_13 | ERR458961
        36.7%
        42%
        1.7
        WT_2 | ERR458878
        31.7%
        42%
        1.5
        WT_2 | ERR458879
        31.1%
        42%
        1.5
        WT_2 | ERR458880
        30.8%
        42%
        1.4
        WT_2 | ERR458881
        30.9%
        42%
        1.3
        WT_2 | ERR458882
        30.2%
        42%
        1.1
        WT_2 | ERR458883
        29.2%
        42%
        1.1
        WT_2 | ERR458884
        33.9%
        42%
        1.5
        WT_21 | ERR459011
        32.4%
        41%
        1.3
        WT_21 | ERR459012
        31.9%
        41%
        1.3
        WT_21 | ERR459013
        31.6%
        41%
        1.3
        WT_21 | ERR459014
        31.1%
        41%
        1.2
        WT_21 | ERR459015
        30.7%
        41%
        1.0
        WT_21 | ERR459016
        30.1%
        41%
        1.0
        WT_21 | ERR459017
        34.5%
        41%
        1.3
        WT_25 | ERR459039
        34.2%
        41%
        1.5
        WT_25 | ERR459040
        33.7%
        41%
        1.5
        WT_25 | ERR459041
        33.7%
        41%
        1.5
        WT_25 | ERR459042
        33.0%
        41%
        1.4
        WT_25 | ERR459043
        32.8%
        41%
        1.2
        WT_25 | ERR459044
        31.5%
        41%
        1.2
        WT_25 | ERR459045
        36.3%
        41%
        1.5
        WT_28 | ERR459060
        33.5%
        42%
        1.5
        WT_28 | ERR459061
        32.6%
        42%
        1.5
        WT_28 | ERR459062
        32.5%
        42%
        1.5
        WT_28 | ERR459063
        32.2%
        42%
        1.3
        WT_28 | ERR459064
        31.8%
        42%
        1.1
        WT_28 | ERR459065
        30.9%
        42%
        1.2
        WT_28 | ERR459066
        35.5%
        42%
        1.5
        WT_5 | ERR458899
        34.6%
        42%
        1.5
        WT_5 | ERR458900
        34.0%
        42%
        1.5
        WT_5 | ERR458901
        33.7%
        42%
        1.5
        WT_5 | ERR458902
        33.9%
        42%
        1.4
        WT_5 | ERR458903
        33.1%
        42%
        1.2
        WT_5 | ERR458904
        31.9%
        42%
        1.2
        WT_5 | ERR458905
        36.8%
        42%
        1.5
        WT_6 | ERR458906
        39.5%
        42%
        2.7
        WT_6 | ERR458907
        38.9%
        42%
        2.7
        WT_6 | ERR458908
        39.6%
        42%
        2.7
        WT_6 | ERR458909
        38.4%
        42%
        2.4
        WT_6 | ERR458910
        37.5%
        42%
        2.1
        WT_6 | ERR458911
        36.7%
        42%
        2.1
        WT_6 | ERR458912
        42.5%
        42%
        2.7

        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 (51bp).

        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.

        112 samples had less than 1% of reads made up of overrepresented sequences

        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..