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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 2021-05-10, 03:55 based on data in: /scratch/gencore/logs/html/HGJM2BGXJ/merged


        General Statistics

        Showing 530 samples.

        loading..

        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 265/265 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        24281706
        5.1
        ReSeq_1408_d02_rep1
        1598154
        0.3
        ReSeq_1408_d04_rep1
        1712026
        0.4
        ReSeq_1408_d06_rep1
        1972542
        0.4
        ReSeq_1408_d08_rep1
        2311680
        0.5
        ReSeq_1408_d12_rep1
        2081945
        0.4
        ReSeq_W17_pDZ_rep1
        1909852
        0.4
        ReSeq_1787_d04_rep1
        83695
        0.0
        ReSeq_1794_d06_rep1
        1733084
        0.4
        ReSeq_1409_d02_rep1
        1582740
        0.3
        ReSeq_1409_d04_rep1
        1904128
        0.4
        ReSeq_1409_d06_rep1
        2113414
        0.4
        ReSeq_1409_d08_rep1
        2365404
        0.5
        ReSeq_1409_d12_rep1
        2190049
        0.5
        ReSeq_W17_Neg_rep1
        251399
        0.1
        ReSeq_1789_d04_rep1
        2168922
        0.5
        ReSeq_1793_d06_rep1
        2090034
        0.4
        ReSeq_1410_d02_rep1
        1833001
        0.4
        ReSeq_1410_d04_rep1
        1206731
        0.3
        ReSeq_1410_d06_rep1
        1820081
        0.4
        ReSeq_1410_d08_rep1
        1468929
        0.3
        ReSeq_1411_d12_rep1
        1522520
        0.3
        ReSeq_W17_HK1073_rep1
        1945406
        0.4
        ReSeq_1793_d04_rep1
        1712183
        0.4
        ReSeq_1789_d06_rep1
        1777288
        0.4
        ReSeq_1411_d02_rep1
        2070495
        0.4
        ReSeq_1411_d04_rep1
        1149329
        0.2
        ReSeq_1411_d06_rep1
        1285672
        0.3
        ReSeq_1411_d08_rep1
        2514880
        0.5
        ReSeq_1414_d12_rep1
        881653
        0.2
        ReSeq_1787_d02_rep1
        2741482
        0.6
        ReSeq_1794_d04_rep1
        2519939
        0.5
        ReSeq2_1787_d06_rep1
        2189337
        0.5
        ReSeq_1412_d02_rep1
        2245950
        0.5
        ReSeq_1412_d04_rep1
        1688804
        0.4
        ReSeq_1412_d06_rep1
        1532836
        0.3
        ReSeq_1412_d08_rep1
        1864053
        0.4
        ReSeq_1415_d12_rep1
        1955574
        0.4
        ReSeq_1789_d02_rep1
        2167017
        0.5
        ReSeq_1797_d04_rep1
        1896657
        0.4
        ReSeq_1803_d06_rep1
        1776916
        0.4
        ReSeq_1413_d02_rep1
        3137
        0.0
        ReSeq_1413_d04_rep1
        2243793
        0.5
        ReSeq_1413_d06_rep1
        1985557
        0.4
        ReSeq_1413_d08_rep1
        2222148
        0.5
        ReSeq_1417_d12_rep1
        2276134
        0.5
        ReSeq_1793_d02_rep1
        2117505
        0.4
        ReSeq_1800_d04_rep1
        2160737
        0.5
        ReSeq_1802_d06_rep1
        2045690
        0.4
        ReSeq_1414_d02_rep1
        1407994
        0.3
        ReSeq_1414_d04_rep1
        1788084
        0.4
        ReSeq_1414_d06_rep1
        1571496
        0.3
        ReSeq_1414_d08_rep1
        1917391
        0.4
        ReSeq_1408_14_rep1
        1456570
        0.3
        ReSeq_1794_d02_rep1
        2165113
        0.5
        ReSeq_1801_d04_rep1
        1735017
        0.4
        ReSeq_1801_d06_rep1
        1854882
        0.4
        ReSeq_1415_d02_rep1
        2217264
        0.5
        ReSeq_1415_d04_rep1
        2079775
        0.4
        ReSeq_1415_d06_rep1
        2076201
        0.4
        ReSeq_1415_d08_rep1
        2672455
        0.6
        ReSeq_1409_14_rep1
        1907258
        0.4
        ReSeq_1797_d02_rep1
        2299582
        0.5
        ReSeq_1802_d04_rep1
        2125279
        0.4
        ReSeq_1800_d06_rep1
        2009799
        0.4
        ReSeq_1416_d02_rep1
        1731300
        0.4
        ReSeq_1416_d04_rep1
        1504580
        0.3
        ReSeq_1416_d06_rep1
        1693713
        0.4
        ReSeq_1416_d08_rep1
        1999060
        0.4
        ReSeq_1411_14_rep1
        2284533
        0.5
        ReSeq_1800_d02_rep1
        1862379
        0.4
        ReSeq_1803_d04_rep1
        1706015
        0.4
        ReSeq_1797_d06_rep1
        1391394
        0.3
        ReSeq_1417_d02_rep1
        2050543
        0.4
        ReSeq_1417_d04_rep1
        1555529
        0.3
        ReSeq_1417_d06_rep1
        2095359
        0.4
        ReSeq_1417_d08_rep1
        2285295
        0.5
        ReSeq_1414_14_rep1
        3084353
        0.6
        ReSeq_1801_d02_rep1
        2634088
        0.5
        ReSeq_1787_d06_rep1
        2028809
        0.4
        ReSeq_1797_d08_rep1
        1541064
        0.3
        ReSeq_1418_d02_rep1
        1988202
        0.4
        ReSeq_1418_d04_rep1
        1925378
        0.4
        ReSeq_1418_d06_rep1
        1955345
        0.4
        ReSeq_1418_d08_rep1
        2248928
        0.5
        ReSeq_1415_14_rep1
        2143749
        0.4
        ReSeq_1803_d02_rep1
        2023564
        0.4
        ReSeq2_1789_d06_rep1
        2099513
        0.4
        ReSeq_1803_d08_rep1
        1743158
        0.4
        ReSeq_1419_d02_rep1
        1719234
        0.4
        ReSeq_1419_d04_rep1
        1365557
        0.3
        ReSeq_1419_d06_rep1
        1858279
        0.4
        ReSeq_1419_d08_rep1
        2225866
        0.5
        ReSeq_1417_14_rep1
        1748451
        0.4
        ReSeq_1802_d02_rep1
        2149156
        0.4
        ReSeq2_1793_d06_rep1
        1630666
        0.3
        ReSeq_1789_d10_rep1
        1257522
        0.3
        ReSeq_1789_d12_rep1
        1929412
        0.4
        ReSeq_1969_d02_rep1
        1580156
        0.3
        ReSeq_1986_d02_rep1
        1802953
        0.4
        ReSeq_1976_d04_rep1
        1943875
        0.4
        ReSeq_1967_d06_rep1
        1810460
        0.4
        ReSeq_1912_d08_rep1
        1853030
        0.4
        ReSeq_1914_d10_rep1
        1434324
        0.3
        ReSeq_1981_d12_rep1
        2022651
        0.4
        ReSeq_1797_d12_rep1
        1962344
        0.4
        ReSeq_1970_d02_rep1
        1708965
        0.4
        ReSeq_1910_d04_rep1
        1757072
        0.4
        ReSeq_1977_d04_rep1
        1821265
        0.4
        ReSeq_1970_d06_rep1
        1944029
        0.4
        ReSeq_1913_d08_rep1
        1885229
        0.4
        ReSeq_1970_d10_rep1
        136.0
        0.0
        ReSeq_Sm18_pDZ_rep1
        1756338
        0.4
        ReSeq_F17_pDZ_rep1
        1531969
        0.3
        ReSeq_1971_d02_rep1
        505639
        0.1
        ReSeq_1911_d04_rep1
        1178398
        0.2
        ReSeq_1980_d04_rep1
        1439624
        0.3
        ReSeq_1971_d06_rep1
        1116378
        0.2
        ReSeq_1914_d08_rep1
        1586719
        0.3
        ReSeq_1971_d10_rep1
        1067540
        0.2
        ReSeq_Sm18_Neg_rep1
        335971
        0.1
        ReSeq_F17_Neg_rep1
        332.0
        0.0
        ReSeq_1972_d02_rep1
        1973262
        0.4
        ReSeq_1912_d04_rep1
        2090775
        0.4
        ReSeq_1981_d04_rep1
        2081526
        0.4
        ReSeq_1973_d06_rep1
        2123288
        0.4
        ReSeq_1970_d08_rep1
        2018328
        0.4
        ReSeq_1976_d10_rep1
        3848524
        0.8
        ReSeq_Sm18_HK1073_rep1
        2764759
        0.6
        ReSeq_F17_HK1073_rep1
        1878444
        0.4
        ReSeq_1973_d02_rep1
        1379017
        0.3
        ReSeq_1913_d04_rep1
        1452981
        0.3
        ReSeq_1983_d04_rep1
        1252694
        0.3
        ReSeq_1974_d06_rep1
        1476186
        0.3
        ReSeq_1971_d08_rep1
        1501820
        0.3
        ReSeq_1981_d10_rep1
        1879805
        0.4
        ReSeq_2234_d02_rep1
        2124446
        0.4
        ReSeq_1910_d02_rep1
        2016143
        0.4
        ReSeq_1974_d02_rep1
        1573621
        0.3
        ReSeq_1914_d04_rep1
        1804899
        0.4
        ReSeq_1984_d04_rep1
        1483308
        0.3
        ReSeq_1976_d06_rep1
        1243563
        0.3
        ReSeq_1976_d08_rep1
        1919602
        0.4
        ReSeq_1983_d10_rep1
        1914001
        0.4
        ReSeq_2232_d02_rep1
        2118699
        0.4
        ReSeq_1913_d02_rep1
        2018653
        0.4
        ReSeq_1976_d02_rep1
        1650885
        0.3
        ReSeq_1915_d04_rep1
        1678426
        0.3
        ReSeq_1986_d04_rep1
        1874469
        0.4
        ReSeq_1977_d06_rep1
        1699064
        0.4
        ReSeq_1981_d08_rep1
        2049429
        0.4
        ReSeq_1984_d10_rep1
        2026870
        0.4
        ReSeq_2251_d02_rep1
        1976935
        0.4
        ReSeq_1914_d02_rep1
        2092231
        0.4
        ReSeq_1977_d02_rep1
        1454826
        0.3
        ReSeq_1967_d04_rep1
        1762301
        0.4
        ReSeq_1910_d06_rep1
        1861204
        0.4
        ReSeq_1980_d06_rep1
        1819566
        0.4
        ReSeq_1983_d08_rep1
        32.0
        0.0
        ReSeq_1986_d10_rep1
        2269311
        0.5
        ReSeq_2238_d02_rep1
        2022289
        0.4
        ReSeq_1915_d02_rep1
        2019049
        0.4
        ReSeq_1980_d02_rep1
        1862805
        0.4
        ReSeq_1970_d04_rep1
        212209
        0.0
        ReSeq_1911_d06_rep1
        1989166
        0.4
        ReSeq_1981_d06_rep1
        1755741
        0.4
        ReSeq_1984_d08_rep1
        1829738
        0.4
        ReSeq_1912_d12_rep1
        2226999
        0.5
        ReSeq_2253_d02_rep1
        2302687
        0.5
        ReSeq_1966_d02_rep1
        1766173
        0.4
        ReSeq_1981_d02_rep1
        1714996
        0.4
        ReSeq_1971_d04_rep1
        1303806
        0.3
        ReSeq_1912_d06_rep1
        1600094
        0.3
        ReSeq_1983_d06_rep1
        1678136
        0.3
        ReSeq_1986_d08_rep1
        1930748
        0.4
        ReSeq_1914_d12_rep1
        1728493
        0.4
        ReSeq_2241_d02_rep1
        1783124
        0.4
        ReSeq_1967_d02_rep1
        1897339
        0.4
        ReSeq_1983_d02_rep1
        1634092
        0.3
        ReSeq_1973_d04_rep1
        1992077
        0.4
        ReSeq_1913_d06_rep1
        2026977
        0.4
        ReSeq_1984_d06_rep1
        2319983
        0.5
        ReSeq_1912_d10_rep1
        2573416
        0.5
        ReSeq_1983_d12_rep1
        1878916
        0.4
        ReSeq_2231_d02_rep1
        1892547
        0.4
        ReSeq_1968_d02_rep1
        1520386
        0.3
        ReSeq_1984_d02_rep1
        1915401
        0.4
        ReSeq_1974_d04_rep1
        1441280
        0.3
        ReSeq_1914_d06_rep1
        1995831
        0.4
        ReSeq_1986_d06_rep1
        2544569
        0.5
        ReSeq_1913_d10_rep1
        2020361
        0.4
        ReSeq_1984_d12_rep1
        1765317
        0.4
        ReSeq_2239_d02_rep1
        1972257
        0.4
        ReSeq_2235_d02_rep1
        1880511
        0.4
        ReSeq_2232_d06_rep1
        1982337
        0.4
        ReSeq_2243_d08_rep1
        2566652
        0.5
        ReSeq_2243_d12_rep1
        2400896
        0.5
        ReSeq_2868_d06_rep1
        2369009
        0.5
        ReSeq_2870_d12_rep1
        2016083
        0.4
        ReSeq_2253_d04_rep1
        1778949
        0.4
        ReSeq_2235_d06_rep1
        13.0
        0.0
        ReSeq_2232_d08_rep1
        1683045
        0.3
        ReSeq_Sp19_pDZ_rep1
        2053102
        0.4
        ReSeq_2867_d06_rep1
        1938984
        0.4
        ReSeq_2868_d12_rep1
        2116186
        0.4
        ReSeq_2241_d04_rep1
        1706284
        0.4
        ReSeq_2251_d06_rep1
        1766049
        0.4
        ReSeq_2238_d08_rep1
        914830
        0.2
        ReSeq_Sp19_Neg_rep1
        24862
        0.0
        ReSeq_2869_d06_rep1
        2334771
        0.5
        ReSeq_2867_d12_rep1
        1479021
        0.3
        ReSeq_2234_d04_rep1
        1828210
        0.4
        ReSeq_2241_d06_rep1
        1896634
        0.4
        ReSeq_2239_d08_rep1
        1730068
        0.4
        ReSeq_Sp19_HK1073_rep1
        2362047
        0.5
        ReSeq_2870_d06_rep1
        2520806
        0.5
        ReSeq_2869_d12_rep1
        1866332
        0.4
        ReSeq_2239_d04_rep1
        1864909
        0.4
        ReSeq_2253_d06_rep1
        1685911
        0.3
        ReSeq2_2243_d08_rep1
        1257928
        0.3
        ReSeq_2867_d02_rep1
        1795452
        0.4
        ReSeq_2867_d08_rep1
        1968662
        0.4
        ReSeq2_1794_d06_rep1
        2297276
        0.5
        ReSeq_2231_d04_rep1
        1319715
        0.3
        ReSeq_2239_d06_rep1
        1461395
        0.3
        ReSeq_2243_d10_rep1
        951642
        0.2
        ReSeq_2869_d02_rep1
        1430354
        0.3
        ReSeq_2870_d08_rep1
        1535049
        0.3
        ReSeq_2233_d02_rep1
        1961834
        0.4
        ReSeq2_2234_d04_rep1
        1994976
        0.4
        ReSeq_2238_d06_rep1
        947573
        0.2
        ReSeq_2239_d10_rep1
        1412842
        0.3
        ReSeq_2868_d02_rep1
        1571072
        0.3
        ReSeq_2869_d08_rep1
        2133465
        0.4
        ReSeq_2239_d12_rep1
        1990468
        0.4
        ReSeq_2233_d04_rep1
        1466947
        0.3
        ReSeq_2231_d06_rep1
        1618903
        0.3
        ReSeq_2253_d10_rep1
        1621695
        0.3
        ReSeq_2870_d02_rep1
        1899693
        0.4
        ReSeq_2868_d08_rep1
        659233
        0.1
        ReSeq_Sp20_pDZ_rep1
        2354256
        0.5
        ReSeq_2232_d04_rep1
        22.0
        0.0
        ReSeq_2234_d06_rep1
        1393329
        0.3
        ReSeq2_2243_d10_rep1
        1763575
        0.4
        ReSeq_2867_d04_rep1
        2279928
        0.5
        ReSeq_2867_d10_rep1
        2011861
        0.4
        ReSeq_Sp20_HK1073_rep1
        2013614
        0.4
        ReSeq_2235_d04_rep1
        1441100
        0.3
        ReSeq_2233_d06_rep1
        55.0
        0.0
        ReSeq_2238_d12_rep1
        78.0
        0.0
        ReSeq_2868_d04_rep1
        58.0
        0.0
        ReSeq_2869_d10_rep1
        81.0
        0.0
        ReSeq_Sp20_Neg_rep1
        96.0
        0.0
        ReSeq_2238_d04_rep1
        1614519
        0.3
        ReSeq_2231_d08_rep1
        1388052
        0.3
        ReSeq_2253_d12_rep1
        1616984
        0.3
        ReSeq_2870_d04_rep1
        1996045
        0.4
        ReSeq_2868_d10_rep1
        1547059
        0.3
        ReSeq_Blank_Well1_rep1
        71.0
        0.0
        ReSeq_2251_d04_rep1
        1484285
        0.3
        ReSeq_2253_d08_rep1
        1099565
        0.2
        ReSeq2_2238_d12_rep1
        1992138
        0.4
        ReSeq_2869_d04_rep1
        1840975
        0.4
        ReSeq_2870_d10_rep1
        1639370
        0.3
        ReSeq_Blank_Well2_rep1
        59.0
        0.0

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        Number of LanesTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        4.0
        545820704
        480772504
        5.0
        2.8

        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 (%)
        GGGGGGGGAGATCTCG
        12951247.0
        53.3
        GGGGGGGGGGGGGGGG
        361806.0
        1.5
        GGGGGGGGAGGCTTAG
        301773.0
        1.2
        GGGGGGGGTATGCAGT
        282274.0
        1.2
        GGGGGGGGCGGAGAGA
        279976.0
        1.1
        GGGGGGGGATAGAGAG
        279974.0
        1.1
        GGGGGGGGAGAGGATA
        270937.0
        1.1
        GGGGGGGGTACTCCTT
        268512.0
        1.1
        GGGGGGGGCTCCTTAC
        257522.0
        1.1
        GGGGGGGGATTAGACG
        225309.0
        0.9
        GGGGGGGGAGCTCTCG
        160295.0
        0.7
        GGGGGGGGCTTAATAG
        155592.0
        0.6
        GGGGGGGGCTAGTCGA
        132850.0
        0.6
        GGGGGGGGACTCTAGG
        99858.0
        0.4
        GGGGGGGGAGCTAGAA
        98098.0
        0.4
        GGGGGGGGTCTTACGC
        92419.0
        0.4
        GGGGGGGGATAGCCTT
        90320.0
        0.4
        TCCTGAGCGGGGGGGG
        33387.0
        0.1
        CGAGGCTGGGGGGGGG
        31426.0
        0.1
        TAAGGCGAGGGGGGGG
        30667.0
        0.1

        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.

        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 (151bp).

        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

        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.

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


        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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