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        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 1.18

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        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 2025-06-04, 15:59 CDT based on data in: /scratch/g/akwitek/wdemos/GSE199976


        General Statistics

        Showing 328/328 rows and 6/9 columns.
        Sample Name% Alignable, M% AlignedM Aligned% Dups% GCM Seqs
        GSM6000724
        40.6%
        GSM6000724_1
        63.0%
        45%
        110.5
        GSM6000724_2
        50.0%
        45%
        110.5
        GSM6000724_STAR
        55.7%
        61.6
        GSM6000725
        46.5%
        GSM6000725_1
        70.8%
        46%
        114.3
        GSM6000725_2
        65.1%
        46%
        114.3
        GSM6000725_STAR
        55.7%
        63.7
        GSM6000726
        44.8%
        GSM6000726_1
        65.2%
        46%
        108.5
        GSM6000726_2
        63.9%
        46%
        108.5
        GSM6000726_STAR
        55.4%
        60.1
        GSM6000727
        38.3%
        GSM6000727_1
        74.6%
        46%
        127.5
        GSM6000727_2
        67.7%
        46%
        127.5
        GSM6000727_STAR
        48.5%
        61.8
        GSM6000728
        47.4%
        GSM6000728_1
        78.5%
        45%
        129.0
        GSM6000728_2
        72.6%
        45%
        129.0
        GSM6000728_STAR
        53.9%
        69.6
        GSM6000729
        30.7%
        GSM6000729_1
        73.4%
        47%
        103.8
        GSM6000729_2
        66.7%
        47%
        103.8
        GSM6000729_STAR
        39.1%
        40.6
        GSM6000730
        36.1%
        GSM6000730_1
        66.0%
        47%
        105.4
        GSM6000730_2
        66.1%
        47%
        105.4
        GSM6000730_STAR
        48.4%
        51.1
        GSM6000731
        38.8%
        GSM6000731_1
        73.6%
        44%
        138.0
        GSM6000731_2
        66.6%
        44%
        138.0
        GSM6000731_STAR
        55.0%
        75.8
        GSM6000732
        44.1%
        GSM6000732_1
        65.2%
        45%
        102.5
        GSM6000732_2
        51.9%
        45%
        102.5
        GSM6000732_STAR
        57.3%
        58.8
        GSM6000733
        28.0%
        GSM6000733_1
        71.1%
        45%
        105.6
        GSM6000733_2
        64.5%
        46%
        105.6
        GSM6000733_STAR
        43.0%
        45.4
        GSM6000734
        32.9%
        GSM6000734_1
        74.3%
        46%
        103.8
        GSM6000734_2
        69.8%
        46%
        103.8
        GSM6000734_STAR
        47.0%
        48.8
        GSM6000735
        44.1%
        GSM6000735_1
        73.3%
        45%
        123.9
        GSM6000735_2
        69.0%
        45%
        123.9
        GSM6000735_STAR
        53.8%
        66.6
        GSM6000736
        50.5%
        GSM6000736_1
        69.1%
        47%
        129.8
        GSM6000736_2
        62.0%
        47%
        129.8
        GSM6000736_STAR
        59.4%
        77.0
        GSM6000737
        33.8%
        GSM6000737_1
        70.2%
        47%
        131.0
        GSM6000737_2
        62.7%
        47%
        131.0
        GSM6000737_STAR
        47.1%
        61.7
        GSM6000738
        46.3%
        GSM6000738_1
        76.2%
        45%
        102.9
        GSM6000738_2
        72.5%
        45%
        102.9
        GSM6000738_STAR
        57.5%
        59.2
        GSM6000739
        38.6%
        GSM6000739_1
        91.6%
        46%
        130.7
        GSM6000739_2
        88.5%
        46%
        130.7
        GSM6000739_STAR
        52.1%
        68.1
        GSM6000740
        42.8%
        GSM6000740_1
        76.0%
        47%
        160.4
        GSM6000740_2
        68.8%
        48%
        160.4
        GSM6000740_STAR
        45.1%
        72.4
        GSM6000741
        52.2%
        GSM6000741_1
        74.6%
        45%
        96.8
        GSM6000741_2
        71.4%
        45%
        96.8
        GSM6000741_STAR
        60.5%
        58.6
        GSM6000742
        44.2%
        GSM6000742_1
        70.9%
        47%
        94.9
        GSM6000742_2
        70.8%
        47%
        94.9
        GSM6000742_STAR
        51.6%
        48.9
        GSM6000743
        39.6%
        GSM6000743_1
        71.7%
        46%
        129.4
        GSM6000743_2
        65.4%
        47%
        129.4
        GSM6000743_STAR
        49.9%
        64.6
        GSM6000744
        51.7%
        GSM6000744_1
        88.1%
        50%
        181.5
        GSM6000744_2
        86.0%
        50%
        181.5
        GSM6000744_STAR
        33.1%
        60.0
        GSM6000745
        34.8%
        GSM6000745_1
        78.3%
        46%
        86.1
        GSM6000745_2
        73.7%
        46%
        86.1
        GSM6000745_STAR
        49.0%
        42.2
        GSM6000746
        31.8%
        GSM6000746_1
        66.8%
        46%
        98.6
        GSM6000746_2
        61.2%
        46%
        98.6
        GSM6000746_STAR
        48.4%
        47.7
        GSM6000747
        42.3%
        GSM6000747_1
        74.1%
        43%
        136.2
        GSM6000747_2
        70.3%
        43%
        136.2
        GSM6000747_STAR
        58.3%
        79.5
        GSM6000748
        38.7%
        GSM6000748_1
        69.7%
        44%
        119.2
        GSM6000748_2
        61.7%
        45%
        119.2
        GSM6000748_STAR
        56.6%
        67.5
        GSM6000749
        52.7%
        GSM6000749_1
        70.5%
        47%
        85.1
        GSM6000749_2
        68.3%
        47%
        85.1
        GSM6000749_STAR
        60.2%
        51.3
        GSM6000750
        38.7%
        GSM6000750_1
        77.0%
        49%
        169.4
        GSM6000750_2
        70.0%
        49%
        169.4
        GSM6000750_STAR
        38.1%
        64.6
        GSM6000751
        40.0%
        GSM6000751_1
        72.3%
        46%
        101.1
        GSM6000751_2
        67.0%
        46%
        101.1
        GSM6000751_STAR
        53.9%
        54.4
        GSM6000752
        40.6%
        GSM6000752_1
        74.0%
        47%
        111.2
        GSM6000752_2
        69.8%
        48%
        111.2
        GSM6000752_STAR
        47.8%
        53.1
        GSM6000753
        42.7%
        GSM6000753_1
        69.5%
        47%
        105.0
        GSM6000753_2
        69.9%
        47%
        105.0
        GSM6000753_STAR
        46.1%
        48.4
        GSM6000754
        53.0%
        GSM6000754_1
        69.8%
        46%
        123.5
        GSM6000754_2
        64.3%
        46%
        123.5
        GSM6000754_STAR
        63.5%
        78.4
        GSM6000755
        41.7%
        GSM6000755_1
        69.1%
        47%
        80.4
        GSM6000755_2
        59.4%
        47%
        80.4
        GSM6000755_STAR
        53.1%
        42.7
        GSM6000756
        35.6%
        GSM6000756_1
        64.5%
        48%
        107.3
        GSM6000756_2
        55.2%
        48%
        107.3
        GSM6000756_STAR
        49.2%
        52.8
        GSM6000757
        33.7%
        GSM6000757_1
        68.8%
        48%
        114.9
        GSM6000757_2
        66.6%
        47%
        114.9
        GSM6000757_STAR
        46.7%
        53.7
        GSM6000758
        55.6%
        GSM6000758_1
        85.8%
        51%
        125.2
        GSM6000758_2
        82.1%
        51%
        125.2
        GSM6000758_STAR
        32.7%
        40.9
        GSM6000759
        37.7%
        GSM6000759_1
        63.8%
        47%
        117.2
        GSM6000759_2
        51.6%
        47%
        117.2
        GSM6000759_STAR
        51.3%
        60.1
        GSM6000760
        34.8%
        GSM6000760_1
        70.9%
        47%
        154.2
        GSM6000760_2
        65.6%
        47%
        154.2
        GSM6000760_STAR
        48.4%
        74.6
        GSM6000761
        41.4%
        GSM6000761_1
        61.9%
        45%
        113.0
        GSM6000761_2
        57.7%
        45%
        113.0
        GSM6000761_STAR
        57.5%
        64.9
        GSM6000762
        30.3%
        GSM6000762_1
        77.1%
        46%
        138.1
        GSM6000762_2
        68.3%
        47%
        138.1
        GSM6000762_STAR
        45.4%
        62.7
        GSM6000763
        41.9%
        GSM6000763_1
        68.4%
        46%
        112.3
        GSM6000763_2
        63.0%
        47%
        112.3
        GSM6000763_STAR
        55.1%
        61.9
        GSM6000764
        31.9%
        GSM6000764_1
        63.7%
        46%
        116.6
        GSM6000764_2
        58.8%
        46%
        116.6
        GSM6000764_STAR
        47.1%
        55.0
        GSM6000765
        26.0%
        GSM6000765_1
        77.5%
        47%
        119.4
        GSM6000765_2
        73.7%
        47%
        119.4
        GSM6000765_STAR
        41.1%
        49.1
        GSM6000766
        38.1%
        GSM6000766_1
        81.0%
        51%
        137.8
        GSM6000766_2
        76.1%
        52%
        137.8
        GSM6000766_STAR
        30.5%
        42.1
        GSM6000767
        39.8%
        GSM6000767_1
        63.2%
        47%
        107.3
        GSM6000767_2
        64.3%
        48%
        107.3
        GSM6000767_STAR
        49.5%
        53.1
        GSM6000768
        43.3%
        GSM6000768_1
        70.2%
        48%
        109.1
        GSM6000768_2
        60.3%
        48%
        109.1
        GSM6000768_STAR
        48.0%
        52.4
        GSM6000769
        37.3%
        GSM6000769_1
        67.5%
        47%
        109.5
        GSM6000769_2
        57.1%
        47%
        109.5
        GSM6000769_STAR
        51.6%
        56.5
        GSM6000770
        34.7%
        GSM6000770_1
        75.0%
        48%
        96.7
        GSM6000770_2
        70.6%
        48%
        96.7
        GSM6000770_STAR
        39.8%
        38.4
        GSM6000771
        29.7%
        GSM6000771_1
        79.2%
        47%
        91.4
        GSM6000771_2
        71.3%
        47%
        91.4
        GSM6000771_STAR
        43.6%
        39.9
        GSM6000772
        80.0%
        GSM6000772_1
        68.0%
        45%
        142.8
        GSM6000772_2
        64.3%
        45%
        142.8
        GSM6000772_STAR
        77.8%
        111.1
        GSM6000773
        42.1%
        GSM6000773_1
        61.1%
        46%
        102.6
        GSM6000773_2
        56.6%
        46%
        102.6
        GSM6000773_STAR
        56.2%
        57.7
        GSM6000774
        33.6%
        GSM6000774_1
        72.5%
        46%
        112.6
        GSM6000774_2
        69.6%
        47%
        112.6
        GSM6000774_STAR
        46.7%
        52.6
        GSM6000775
        36.3%
        GSM6000775_1
        59.2%
        48%
        46.1
        GSM6000775_2
        53.3%
        48%
        46.1
        GSM6000775_STAR
        48.9%
        22.6
        GSM6000776
        28.8%
        GSM6000776_1
        74.4%
        46%
        108.1
        GSM6000776_2
        67.0%
        47%
        108.1
        GSM6000776_STAR
        46.8%
        50.6
        GSM6000777
        41.6%
        GSM6000777_1
        65.6%
        46%
        107.6
        GSM6000777_2
        62.3%
        46%
        107.6
        GSM6000777_STAR
        57.4%
        61.8
        GSM6000778
        41.6%
        GSM6000778_1
        66.8%
        47%
        137.0
        GSM6000778_2
        62.8%
        47%
        137.0
        GSM6000778_STAR
        53.4%
        73.1
        GSM6000779
        31.1%
        GSM6000779_1
        55.3%
        48%
        46.0
        GSM6000779_2
        49.8%
        48%
        46.0
        GSM6000779_STAR
        45.8%
        21.1
        GSM6000780
        32.5%
        GSM6000780_1
        68.9%
        46%
        147.8
        GSM6000780_2
        66.6%
        46%
        147.8
        GSM6000780_STAR
        49.5%
        73.1
        GSM6000781
        42.8%
        GSM6000781_1
        65.5%
        46%
        107.5
        GSM6000781_2
        61.5%
        46%
        107.5
        GSM6000781_STAR
        55.1%
        59.2
        GSM6000782
        34.0%
        GSM6000782_1
        71.2%
        47%
        159.6
        GSM6000782_2
        67.1%
        48%
        159.6
        GSM6000782_STAR
        53.1%
        84.7
        GSM6000783
        46.9%
        GSM6000783_1
        74.7%
        48%
        134.9
        GSM6000783_2
        70.9%
        48%
        134.9
        GSM6000783_STAR
        48.1%
        64.9
        GSM6000784
        36.4%
        GSM6000784_1
        68.5%
        48%
        75.4
        GSM6000784_2
        65.4%
        48%
        75.4
        GSM6000784_STAR
        47.1%
        35.5
        GSM6000785
        26.8%
        GSM6000785_1
        83.8%
        47%
        151.0
        GSM6000785_2
        79.8%
        47%
        151.0
        GSM6000785_STAR
        41.3%
        62.3
        GSM6000786
        63.1%
        GSM6000786_1
        80.6%
        50%
        126.9
        GSM6000786_2
        76.1%
        49%
        126.9
        GSM6000786_STAR
        45.1%
        57.3
        GSM6000787
        37.9%
        GSM6000787_1
        63.0%
        47%
        112.0
        GSM6000787_2
        64.8%
        46%
        112.0
        GSM6000787_STAR
        50.4%
        56.4
        GSM6000788
        36.1%
        GSM6000788_1
        64.2%
        48%
        168.4
        GSM6000788_2
        54.1%
        48%
        168.4
        GSM6000788_STAR
        47.4%
        79.8
        GSM6000789
        34.4%
        GSM6000789_1
        73.0%
        48%
        132.0
        GSM6000789_2
        69.3%
        48%
        132.0
        GSM6000789_STAR
        49.1%
        64.8
        GSM6000790
        45.6%
        GSM6000790_1
        72.1%
        47%
        125.4
        GSM6000790_2
        68.2%
        47%
        125.4
        GSM6000790_STAR
        53.4%
        66.9
        GSM6000791
        31.7%
        GSM6000791_1
        77.0%
        46%
        105.6
        GSM6000791_2
        73.1%
        47%
        105.6
        GSM6000791_STAR
        45.9%
        48.5
        GSM6000792
        34.7%
        GSM6000792_1
        61.1%
        47%
        84.0
        GSM6000792_2
        60.4%
        46%
        84.0
        GSM6000792_STAR
        51.0%
        42.8
        GSM6000793
        35.6%
        GSM6000793_1
        80.8%
        48%
        128.5
        GSM6000793_2
        77.1%
        48%
        128.5
        GSM6000793_STAR
        38.9%
        50.0
        GSM6000794
        42.0%
        GSM6000794_1
        64.1%
        46%
        115.6
        GSM6000794_2
        66.3%
        46%
        115.6
        GSM6000794_STAR
        54.6%
        63.1
        GSM6000795
        40.3%
        GSM6000795_1
        72.1%
        46%
        154.2
        GSM6000795_2
        66.9%
        46%
        154.2
        GSM6000795_STAR
        54.2%
        83.6
        GSM6000796
        51.2%
        GSM6000796_1
        78.4%
        49%
        129.0
        GSM6000796_2
        74.1%
        50%
        129.0
        GSM6000796_STAR
        36.3%
        46.9
        GSM6000797
        44.2%
        GSM6000797_1
        67.0%
        46%
        122.4
        GSM6000797_2
        50.1%
        46%
        122.4
        GSM6000797_STAR
        57.3%
        70.1
        GSM6000798
        27.4%
        GSM6000798_1
        83.0%
        46%
        139.7
        GSM6000798_2
        73.7%
        46%
        139.7
        GSM6000798_STAR
        43.5%
        60.8
        GSM6000799
        37.4%
        GSM6000799_1
        62.1%
        46%
        153.9
        GSM6000799_2
        51.4%
        46%
        153.9
        GSM6000799_STAR
        53.1%
        81.7
        GSM6000800
        37.0%
        GSM6000800_1
        66.5%
        46%
        116.9
        GSM6000800_2
        64.2%
        46%
        116.9
        GSM6000800_STAR
        51.9%
        60.6
        GSM6000801
        37.7%
        GSM6000801_1
        68.7%
        47%
        118.4
        GSM6000801_2
        64.6%
        47%
        118.4
        GSM6000801_STAR
        57.8%
        68.4
        GSM6000802
        50.2%
        GSM6000802_1
        78.3%
        48%
        127.3
        GSM6000802_2
        72.5%
        48%
        127.3
        GSM6000802_STAR
        45.7%
        58.1
        GSM6000803
        31.2%
        GSM6000803_1
        75.6%
        50%
        108.6
        GSM6000803_2
        72.5%
        50%
        108.6
        GSM6000803_STAR
        41.3%
        44.8
        GSM6000804
        49.4%
        GSM6000804_1
        83.7%
        51%
        114.2
        GSM6000804_2
        80.2%
        51%
        114.2
        GSM6000804_STAR
        34.1%
        39.0
        GSM6000805
        49.3%
        GSM6000805_1
        85.3%
        50%
        110.0
        GSM6000805_2
        81.9%
        50%
        110.0
        GSM6000805_STAR
        31.7%
        34.9

        Rsem

        Rsem RSEM (RNA-Seq by Expectation-Maximization) is a software package forestimating gene and isoform expression levels from RNA-Seq data.DOI: 10.1186/1471-2105-12-323.

        Mapped Reads

        A breakdown of how all reads were aligned for each sample.

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        Multimapping rates

        A frequency histogram showing how many reads were aligned to n reference regions.

        In an ideal world, every sequence reads would align uniquely to a single location in the reference. However, due to factors such as repeititve sequences, short reads and sequencing errors, reads can be align to the reference 0, 1 or more times. This plot shows the frequency of each factor of multimapping. Good samples should have the majority of reads aligning once.

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        STAR

        STAR is an ultrafast universal RNA-seq aligner.DOI: 10.1093/bioinformatics/bts635.

        Alignment Scores

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        FastQ Screen

        Version: 0.15.1

        FastQ Screen allows you to screen a library of sequences in FastQ format against a set of sequence databases so you can see if the composition of the library matches with what you expect.DOI: 10.12688/f1000research.15931.2.

        Mapped Reads

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        FastQC

        Versions: 0.11.8, 0.11.9

        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Sequence Length Distribution

        All samples have sequences of a single length (150bp).

        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Overrepresented sequences by sample

        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 overrepresented.

        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 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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        Top overrepresented sequences

        Top overrepresented sequences across all samples. The table shows 20 most overrepresented sequences across all samples, ranked by the number of samples they occur in.

        Showing 20/20 rows and 3/3 columns.
        Overrepresented sequenceSamplesOccurrences% of all reads
        GTGGAATTAGTGTGTGTAAGTATGTATGTTGAGCTTGAACGCTTTCTTTA
        82
        26379884
        0.1361%
        CCTTAGGCAACCTGGTGGTCCCCCGCTCCCGGGAGGTCACCATATTGATG
        82
        27462834
        0.1417%
        GCTACCTTTGCACGGTCAGGATACCGCGGCCGTTTAACTTTAGTCACTGG
        77
        17183574
        0.0887%
        CCCCAACCGAAATTTTTTAGTTCATATTTATTTTGTTTTAGCCCATTAGG
        77
        21839081
        0.1127%
        CTTGAGTCCAGGAGTTCTGGGCTGTAGTGCGCTATGCCGATCGGGTGTCC
        75
        14714507
        0.0759%
        CGTTGATCAATAATTGGGTCAATAAGATATTAGTATTACTTTGACTTGTG
        75
        15399316
        0.0795%
        CCAGGCTGGAGTGCAGTGGCTATTCACAGGCGCGATCCCACTACTGATCA
        73
        18502317
        0.0955%
        GTTTAACTTTAGTCACTGGGCAGGCAATGCCTCTAATACTTGTTATGCTA
        69
        11954714
        0.0617%
        GCTCGTTTGGTTTCGGGGTTCTTAGCTTAAATTCTTTTTGTTAAGGATTT
        67
        14083126
        0.0727%
        GTTCTGGGCTGTAGTGCGCTATGCCGATCGGGTGTCCGCACTAAGTTCGG
        66
        11638031
        0.0601%
        CCCTCCTTAGGCAACCTGGTGGTCCCCCGCTCCCGGGAGGTCACCATATT
        65
        10265802
        0.0530%
        ATTAGGTTGTTTTTATATAAGTTGAACTAGTAAATTGAAGCTCCATAGGG
        65
        13349183
        0.0689%
        GCACGTTTTACGCCGAAAATAATTAGTTTGGGTTAATCGTATGACCGCGG
        64
        12566296
        0.0649%
        CTCCGTTTCCGACCTGGGCCGGTTCACCCCTCCTTAGGCAACCTGGTGGT
        62
        14941307
        0.0771%
        GTCCTTTCGTACTGGGAGAAATTGTAAATAGATAGAAACCGACCTGGATT
        59
        10181892
        0.0525%
        GTTTGGTTTCGGGGTTCTTAGCTTAAATTCTTTTTGTTAAGGATTTTCTA
        58
        10217771
        0.0527%
        GTAGGACTTTAATCGTTGAACAAACGAACCATTAATAGCTTCTGCACCAT
        56
        9563236
        0.0494%
        CAGGCTGGAGTGCAGTGGCTATTCACAGGCGCGATCCCACTACTGATCAG
        56
        9582677
        0.0495%
        CAGGAGGATCGCTTGAGTCCAGGAGTTCTGGGCTGTAGTGCGCTATGCCG
        56
        11162905
        0.0576%
        GTTGGGTTAGTACCTATGATTCGATAATTGACAATGGTTATCCGGGTTGT
        55
        10924563
        0.0564%

        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.

        Flat image plot. Toolbox functions such as highlighting / hiding samples will not work (see the docs).


        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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        Software Versions

        Software Versions lists versions of software tools extracted from file contents.

        SoftwareVersion
        FastQ Screen0.15.1
        FastQC0.11.8, 0.11.9