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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 2021-05-12, 14:19 based on data in: /scratch/gencore/logs/html/HGGF2BGXJ/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
        25233420
        5.3
        ReSeq_1408_d02_rep2
        1952781
        0.4
        ReSeq_1408_d04_rep2
        1733910
        0.4
        ReSeq_1408_d06_rep2
        2004421
        0.4
        ReSeq_1408_d08_rep2
        1547632
        0.3
        ReSeq_1408_d12_rep2
        1884299
        0.4
        ReSeq_W17_pDZ_rep2
        1439909
        0.3
        ReSeq_1787_d04_rep2
        1583851
        0.3
        ReSeq_1794_d06_rep2
        1770952
        0.4
        ReSeq_1409_d02_rep2
        2099475
        0.4
        ReSeq_1409_d04_rep2
        1610276
        0.3
        ReSeq_1409_d06_rep2
        2160724
        0.5
        ReSeq_1409_d08_rep2
        2127482
        0.4
        ReSeq_1409_d12_rep2
        2361042
        0.5
        ReSeq_W17_Neg_rep2
        147629
        0.0
        ReSeq_1789_d04_rep2
        1584198
        0.3
        ReSeq_1793_d06_rep2
        1412984
        0.3
        ReSeq_1410_d02_rep2
        1473038
        0.3
        ReSeq_1410_d04_rep2
        1775708
        0.4
        ReSeq_1410_d06_rep2
        1079233
        0.2
        ReSeq_1410_d08_rep2
        1270860
        0.3
        ReSeq_1411_d12_rep2
        1932624
        0.4
        ReSeq_W17_HK1073_rep2
        948347
        0.2
        ReSeq_1793_d04_rep2
        1262835
        0.3
        ReSeq_1789_d06_rep2
        1573230
        0.3
        ReSeq_1411_d02_rep2
        2629670
        0.6
        ReSeq_1411_d04_rep2
        1917660
        0.4
        ReSeq_1411_d06_rep2
        1691957
        0.4
        ReSeq_1411_d08_rep2
        2241663
        0.5
        ReSeq_1414_d12_rep2
        1093250
        0.2
        ReSeq_1787_d02_rep2
        1575877
        0.3
        ReSeq_1794_d04_rep2
        1641086
        0.3
        ReSeq_1787_d06_rep2
        1831275
        0.4
        ReSeq_1412_d02_rep2
        2239449
        0.5
        ReSeq_1412_d04_rep2
        1517788
        0.3
        ReSeq_1412_d06_rep2
        1678130
        0.4
        ReSeq_1412_d08_rep2
        1983542
        0.4
        ReSeq_1415_d12_rep2
        1519664
        0.3
        ReSeq_1789_d02_rep2
        1448458
        0.3
        ReSeq_1797_d04_rep2
        1878360
        0.4
        ReSeq_1803_d06_rep2
        2338106
        0.5
        ReSeq_1413_d02_rep2
        2186725
        0.5
        ReSeq_1413_d04_rep2
        435423
        0.1
        ReSeq_1413_d06_rep2
        1794760
        0.4
        ReSeq_1413_d08_rep2
        1667954
        0.4
        ReSeq_1417_d12_rep2
        1803855
        0.4
        ReSeq_1793_d02_rep2
        1287682
        0.3
        ReSeq_1800_d04_rep2
        1698091
        0.4
        ReSeq_1802_d06_rep2
        1949631
        0.4
        ReSeq_1414_d02_rep2
        1704959
        0.4
        ReSeq_1414_d04_rep2
        1163070
        0.2
        ReSeq_1414_d06_rep2
        1404646
        0.3
        ReSeq_1414_d08_rep2
        1905412
        0.4
        ReSeq_1408_14_rep2
        1677090
        0.4
        ReSeq_1794_d02_rep2
        1357044
        0.3
        ReSeq_1801_d04_rep2
        1619322
        0.3
        ReSeq_1801_d06_rep2
        1676451
        0.4
        ReSeq_1415_d02_rep2
        2199043
        0.5
        ReSeq_1415_d04_rep2
        1296221
        0.3
        ReSeq_1415_d06_rep2
        1449727
        0.3
        ReSeq_1415_d08_rep2
        1677560
        0.4
        ReSeq_1409_14_rep2
        1669000
        0.4
        ReSeq_1797_d02_rep2
        1129774
        0.2
        ReSeq_1802_d04_rep2
        1771141
        0.4
        ReSeq_1800_d06_rep2
        1883823
        0.4
        ReSeq_1416_d02_rep2
        2024306
        0.4
        ReSeq_1416_d04_rep2
        1936968
        0.4
        ReSeq_1416_d06_rep2
        1908109
        0.4
        ReSeq_1416_d08_rep2
        2367639
        0.5
        ReSeq_1411_14_rep2
        2016716
        0.4
        ReSeq_1800_d02_rep2
        1792369
        0.4
        ReSeq_1803_d04_rep2
        1688774
        0.4
        ReSeq_1797_d06_rep2
        2499908
        0.5
        ReSeq_1417_d02_rep2
        1398722
        0.3
        ReSeq_1417_d04_rep2
        1443122
        0.3
        ReSeq_1417_d06_rep2
        1856407
        0.4
        ReSeq_1417_d08_rep2
        1663235
        0.3
        ReSeq_1414_14_rep2
        1869943
        0.4
        ReSeq_1801_d02_rep2
        1452799
        0.3
        ReSeq2_1787_d06_rep2
        1413782
        0.3
        ReSeq_1797_d08_rep2
        1397317
        0.3
        ReSeq_1418_d02_rep2
        2006681
        0.4
        ReSeq_1418_d04_rep2
        1846802
        0.4
        ReSeq_1418_d06_rep2
        1808241
        0.4
        ReSeq_1418_d08_rep2
        1858805
        0.4
        ReSeq_1415_14_rep2
        1701994
        0.4
        ReSeq_1803_d02_rep2
        1695001
        0.4
        ReSeq2_1789_d06_rep2
        1164478
        0.2
        ReSeq_1803_d08_rep2
        1515538
        0.3
        ReSeq_1419_d02_rep2
        5016
        0.0
        ReSeq_1419_d04_rep2
        1238050
        0.3
        ReSeq_1419_d06_rep2
        1374780
        0.3
        ReSeq_1419_d08_rep2
        1768605
        0.4
        ReSeq_1417_14_rep2
        1195424
        0.3
        ReSeq_1802_d02_rep2
        2015795
        0.4
        ReSeq2_1793_d06_rep2
        1216239
        0.3
        ReSeq_1789_d10_rep2
        1471922
        0.3
        ReSeq_1789_d12_rep2
        976724
        0.2
        ReSeq_1969_d02_rep2
        1285561
        0.3
        ReSeq_1986_d02_rep2
        1192863
        0.3
        ReSeq_1976_d04_rep2
        1561759
        0.3
        ReSeq_1967_d06_rep2
        1446986
        0.3
        ReSeq_1912_d08_rep2
        1631653
        0.3
        ReSeq_1914_d10_rep2
        1048370
        0.2
        ReSeq_1981_d12_rep2
        1471178
        0.3
        ReSeq_1797_d12_rep2
        1315599
        0.3
        ReSeq_1970_d02_rep2
        1254605
        0.3
        ReSeq_1910_d04_rep2
        1189533
        0.2
        ReSeq_1977_d04_rep2
        1829848
        0.4
        ReSeq_1970_d06_rep2
        2078959
        0.4
        ReSeq_1913_d08_rep2
        1492252
        0.3
        ReSeq_1970_d10_rep2
        9037
        0.0
        ReSeq_Sm18_pDZ_rep2
        1604007
        0.3
        ReSeq_F17_pDZ_rep2
        1685934
        0.4
        ReSeq_1971_d02_rep2
        328152
        0.1
        ReSeq_1911_d04_rep2
        1304063
        0.3
        ReSeq_1980_d04_rep2
        1676167
        0.4
        ReSeq_1971_d06_rep2
        1817210
        0.4
        ReSeq_1914_d08_rep2
        1414590
        0.3
        ReSeq_1971_d10_rep2
        1077662
        0.2
        ReSeq_Sm18_Neg_rep2
        138761
        0.0
        ReSeq_F17_Neg_rep2
        29924
        0.0
        ReSeq_1972_d02_rep2
        1449591
        0.3
        ReSeq_1912_d04_rep2
        1543354
        0.3
        ReSeq_1981_d04_rep2
        1593782
        0.3
        ReSeq_1973_d06_rep2
        1877243
        0.4
        ReSeq_1970_d08_rep2
        2190363
        0.5
        ReSeq_1976_d10_rep2
        1761066
        0.4
        ReSeq_Sm18_HK1073_rep2
        1820633
        0.4
        ReSeq_F17_HK1073_rep2
        708850
        0.1
        ReSeq_1973_d02_rep2
        998841
        0.2
        ReSeq_1913_d04_rep2
        1324734
        0.3
        ReSeq_1983_d04_rep2
        1466492
        0.3
        ReSeq_1974_d06_rep2
        1806537
        0.4
        ReSeq_1971_d08_rep2
        1396356
        0.3
        ReSeq_1981_d10_rep2
        1198713
        0.3
        ReSeq_2234_d02_rep2
        1469120
        0.3
        ReSeq_1910_d02_rep2
        1168337
        0.2
        ReSeq_1974_d02_rep2
        1077370
        0.2
        ReSeq_1914_d04_rep2
        1434648
        0.3
        ReSeq_1984_d04_rep2
        1204869
        0.3
        ReSeq_1976_d06_rep2
        718602
        0.2
        ReSeq_1976_d08_rep2
        890506
        0.2
        ReSeq_1983_d10_rep2
        1158094
        0.2
        ReSeq_2232_d02_rep2
        1597965
        0.3
        ReSeq_1913_d02_rep2
        1392198
        0.3
        ReSeq_1976_d02_rep2
        1152989
        0.2
        ReSeq_1915_d04_rep2
        1827899
        0.4
        ReSeq_1986_d04_rep2
        1783813
        0.4
        ReSeq_1977_d06_rep2
        1191115
        0.3
        ReSeq_1981_d08_rep2
        2201052
        0.5
        ReSeq_1984_d10_rep2
        815373
        0.2
        ReSeq_2251_d02_rep2
        1827212
        0.4
        ReSeq_1914_d02_rep2
        581273
        0.1
        ReSeq_1977_d02_rep2
        1073599
        0.2
        ReSeq_1967_d04_rep2
        911988
        0.2
        ReSeq_1910_d06_rep2
        1318324
        0.3
        ReSeq_1980_d06_rep2
        812644
        0.2
        ReSeq_1983_d08_rep2
        1145686
        0.2
        ReSeq_1986_d10_rep2
        1370924
        0.3
        ReSeq_2238_d02_rep2
        1372526
        0.3
        ReSeq_1915_d02_rep2
        1207222
        0.3
        ReSeq_1980_d02_rep2
        1229190
        0.3
        ReSeq_1970_d04_rep2
        145029
        0.0
        ReSeq_1911_d06_rep2
        1667040
        0.4
        ReSeq_1981_d06_rep2
        1434433
        0.3
        ReSeq_1984_d08_rep2
        1372668
        0.3
        ReSeq_1912_d12_rep2
        2321732
        0.5
        ReSeq_2253_d02_rep2
        2285909
        0.5
        ReSeq_1966_d02_rep2
        883237
        0.2
        ReSeq_1981_d02_rep2
        820188
        0.2
        ReSeq_1971_d04_rep2
        96489
        0.0
        ReSeq_1912_d06_rep2
        1353729
        0.3
        ReSeq_1983_d06_rep2
        1062561
        0.2
        ReSeq_1986_d08_rep2
        1082667
        0.2
        ReSeq_1914_d12_rep2
        530377
        0.1
        ReSeq_2241_d02_rep2
        708167
        0.1
        ReSeq_1967_d02_rep2
        1073977
        0.2
        ReSeq_1983_d02_rep2
        1446767
        0.3
        ReSeq_1973_d04_rep2
        1210636
        0.3
        ReSeq_1913_d06_rep2
        1879541
        0.4
        ReSeq_1984_d06_rep2
        1604465
        0.3
        ReSeq_1912_d10_rep2
        1965117
        0.4
        ReSeq_1983_d12_rep2
        1767855
        0.4
        ReSeq_2231_d02_rep2
        1800340
        0.4
        ReSeq_1968_d02_rep2
        1036310
        0.2
        ReSeq_1984_d02_rep2
        943806
        0.2
        ReSeq_1974_d04_rep2
        1456410
        0.3
        ReSeq_1914_d06_rep2
        1405975
        0.3
        ReSeq_1986_d06_rep2
        1719877
        0.4
        ReSeq_1913_d10_rep2
        1171744
        0.2
        ReSeq_1984_d12_rep2
        1122363
        0.2
        ReSeq_2239_d02_rep2
        1078867
        0.2
        ReSeq_2235_d02_rep2
        2409286
        0.5
        ReSeq_2232_d06_rep2
        3013254
        0.6
        ReSeq_2243_d08_rep2
        6589587
        1.4
        ReSeq_2243_d12_rep2
        2505689
        0.5
        ReSeq_2868_d06_rep2
        1982861
        0.4
        ReSeq_2870_d12_rep2
        2044028
        0.4
        ReSeq_2253_d04_rep2
        2171031
        0.5
        ReSeq_2235_d06_rep2
        2180107
        0.5
        ReSeq_2232_d08_rep2
        2458401
        0.5
        ReSeq_Sp19_pDZ_rep2
        2756250
        0.6
        ReSeq_2867_d06_rep2
        3749677
        0.8
        ReSeq_2868_d12_rep2
        2059078
        0.4
        ReSeq_2241_d04_rep2
        1443871
        0.3
        ReSeq_2251_d06_rep2
        2163999
        0.5
        ReSeq_2238_d08_rep2
        1802686
        0.4
        ReSeq_Sp19_Neg_rep2
        57771
        0.0
        ReSeq_2869_d06_rep2
        3327327
        0.7
        ReSeq_2867_d12_rep2
        2223401
        0.5
        ReSeq_2234_d04_rep2
        2426067
        0.5
        ReSeq_2241_d06_rep2
        2340249
        0.5
        ReSeq_2239_d08_rep2
        1828880
        0.4
        ReSeq_Sp19_HK1073_rep2
        2678113
        0.6
        ReSeq_2870_d06_rep2
        5273782
        1.1
        ReSeq_2869_d12_rep2
        1572982
        0.3
        ReSeq_2239_d04_rep2
        2090897
        0.4
        ReSeq_2253_d06_rep2
        2593384
        0.5
        ReSeq2_2243_d08_rep2
        2521317
        0.5
        ReSeq_2867_d02_rep2
        2105665
        0.4
        ReSeq_2867_d08_rep2
        3038899
        0.6
        ReSeq2_1794_d06_rep2
        603021
        0.1
        ReSeq_2231_d04_rep2
        3966320
        0.8
        ReSeq_2239_d06_rep2
        3166680
        0.7
        ReSeq_2243_d10_rep2
        1847906
        0.4
        ReSeq_2869_d02_rep2
        2877555
        0.6
        ReSeq_2870_d08_rep2
        2495688
        0.5
        ReSeq_2233_d02_rep2
        1488211
        0.3
        ReSeq2_2234_d04_rep2
        2178619
        0.5
        ReSeq_2238_d06_rep2
        2385787
        0.5
        ReSeq_2239_d10_rep2
        1993340
        0.4
        ReSeq_2868_d02_rep2
        1742849
        0.4
        ReSeq_2869_d08_rep2
        2806109
        0.6
        ReSeq_2239_d12_rep2
        2509444
        0.5
        ReSeq_2233_d04_rep2
        2764188
        0.6
        ReSeq_2231_d06_rep2
        2526664
        0.5
        ReSeq_2253_d10_rep2
        2509952
        0.5
        ReSeq_2870_d02_rep2
        1854358
        0.4
        ReSeq_2868_d08_rep2
        1459335
        0.3
        ReSeq_Sp20_pDZ_rep2
        1664655
        0.3
        ReSeq_2232_d04_rep2
        2904101
        0.6
        ReSeq_2234_d06_rep2
        2632372
        0.6
        ReSeq2_2243_d10_rep2
        2529852
        0.5
        ReSeq_2867_d04_rep2
        2515194
        0.5
        ReSeq_2867_d10_rep2
        2300312
        0.5
        ReSeq_Sp20_HK1073_rep2
        2143041
        0.5
        ReSeq_2235_d04_rep2
        1543871
        0.3
        ReSeq_2233_d06_rep2
        2015419
        0.4
        ReSeq_2238_d12_rep2
        1622512
        0.3
        ReSeq_2868_d04_rep2
        214841
        0.0
        ReSeq_2869_d10_rep2
        1392130
        0.3
        ReSeq_Sp20_Neg_rep2
        112702
        0.0
        ReSeq_2238_d04_rep2
        2398202
        0.5
        ReSeq_2231_d08_rep2
        2340279
        0.5
        ReSeq_2253_d12_rep2
        3866552
        0.8
        ReSeq_2870_d04_rep2
        2223647
        0.5
        ReSeq_2868_d10_rep2
        1805352
        0.4
        ReSeq_Blank_Well1_rep2
        76.0
        0.0
        ReSeq_2251_d04_rep2
        2628657
        0.6
        ReSeq_2253_d08_rep2
        2060305
        0.4
        ReSeq2_2238_d12_rep2
        4754325
        1.0
        ReSeq_2869_d04_rep2
        3108737
        0.7
        ReSeq_2870_d10_rep2
        3214775
        0.7
        ReSeq_Blank_Well2_rep2
        80.0
        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
        565113344
        475176006
        5.3
        2.9

        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
        13435105.0
        53.2
        GGGGGGGGGGGGGGGG
        546301.0
        2.2
        GGGGGGGGTATGCAGT
        271997.0
        1.1
        GGGGGGGGCTCCTTAC
        244622.0
        1.0
        GGGGGGGGATAGAGAG
        240488.0
        0.9
        GGGGGGGGTACTCCTT
        239141.0
        0.9
        GGGGGGGGCGGAGAGA
        234206.0
        0.9
        GGGGGGGGAGGCTTAG
        212186.0
        0.8
        GGGGGGGGAGAGGATA
        209832.0
        0.8
        GGGGGGGGATTAGACG
        190717.0
        0.8
        GGGGGGGGAGCTCTCG
        180386.0
        0.7
        GGGGGGGGCTTAATAG
        149330.0
        0.6
        GGGGGGGGAGCTAGAA
        146591.0
        0.6
        GGGGGGGGACTCTAGG
        142511.0
        0.6
        GGGGGGGGCTAGTCGA
        121984.0
        0.5
        GGGGGGGGTCTTACGC
        86223.0
        0.3
        GGGGGGGGATAGCCTT
        61969.0
        0.2
        NNNNNNNNNNNNNNNN
        53802.0
        0.2
        ACTCGCTAGGGGGGGG
        26041.0
        0.1
        GGGGGGGGAGATATCG
        25432.0
        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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