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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 2024-05-11, 07:29 based on data in: /vast/gencore/GENEFLOW/work/1b/dec7900e64551f3fc27fb1692a01d5/1


        General Statistics

        Showing 518 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 259/259 rows and 2/2 columns.
        LibraryTotal Read CountPortion (%)
        undetermined_library
        245504
        20.8
        A_thaliana_001
        2968
        0.3
        A_thaliana_002
        3287
        0.3
        A_thaliana_003
        3929
        0.3
        A_thaliana_004
        919.0
        0.1
        A_thaliana_005
        4255
        0.4
        A_thaliana_006
        3884
        0.3
        A_thaliana_007
        4239
        0.4
        A_thaliana_008
        2689
        0.2
        A_thaliana_009
        3464
        0.3
        A_thaliana_010
        4306
        0.4
        A_thaliana_011
        4876
        0.4
        A_thaliana_012
        3423
        0.3
        A_thaliana_013
        3832
        0.3
        A_thaliana_014
        3427
        0.3
        A_thaliana_015
        5385
        0.5
        A_thaliana_016
        3782
        0.3
        ath_dve_017
        3619
        0.3
        A_thaliana_018
        4947
        0.4
        A_thaliana_019
        5177
        0.4
        A_thaliana_020
        5967
        0.5
        ath_dve_021
        2327
        0.2
        A_thaliana_022
        3541
        0.3
        A_thaliana_023
        6838
        0.6
        A_thaliana_024
        4522
        0.4
        A_thaliana_025
        0.0
        0.0
        A_thaliana_026
        0.0
        0.0
        A_thaliana_027
        0.0
        0.0
        A_thaliana_028
        0.0
        0.0
        A_thaliana_029
        0.0
        0.0
        A_thaliana_030
        0.0
        0.0
        A_thaliana_031
        0.0
        0.0
        A_thaliana_032
        2192
        0.2
        ath_dve_033
        2227
        0.2
        A_thaliana_034
        3588
        0.3
        A_thaliana_035
        2602
        0.2
        A_thaliana_036
        3733
        0.3
        A_thaliana_037
        3796
        0.3
        A_thaliana_038
        3475
        0.3
        A_thaliana_039
        4676
        0.4
        A_thaliana_040
        2254
        0.2
        A_thaliana_041
        2987
        0.3
        A_thaliana_042
        1531
        0.1
        A_thaliana_043
        2960
        0.3
        A_thaliana_044
        2504
        0.2
        A_thaliana_045
        2284
        0.2
        A_thaliana_046
        2762
        0.2
        A_thaliana_047
        2436
        0.2
        A_thaliana_048
        2193
        0.2
        A_thaliana_049
        5278
        0.4
        A_thaliana_050
        2473
        0.2
        A_thaliana_051
        1665
        0.1
        A_thaliana_052
        1796
        0.2
        A_thaliana_053
        2248
        0.2
        A_thaliana_054
        2357
        0.2
        A_thaliana_055
        280.0
        0.0
        A_thaliana_056
        4388
        0.4
        A_thaliana_057
        3207
        0.3
        A_thaliana_058
        5161
        0.4
        A_thaliana_059
        5898
        0.5
        A_thaliana_060
        1312
        0.1
        A_thaliana_061
        6507
        0.6
        A_thaliana_062
        5741
        0.5
        A_thaliana_063
        6625
        0.6
        A_thaliana_064
        0.0
        0.0
        A_thaliana_065
        0.0
        0.0
        A_thaliana_066
        0.0
        0.0
        A_thaliana_067
        0.0
        0.0
        A_thaliana_068
        0.0
        0.0
        A_thaliana_069
        0.0
        0.0
        A_thaliana_070
        0.0
        0.0
        A_thaliana_071
        0.0
        0.0
        A_thaliana_072
        2214
        0.2
        A_thaliana_073
        8718
        0.7
        A_thaliana_074
        420.0
        0.0
        A_thaliana_075
        6412
        0.5
        A_thaliana_076
        6434
        0.5
        A_thaliana_077
        3334
        0.3
        A_thaliana_078
        2060
        0.2
        A_thaliana_079
        2653
        0.2
        A_thaliana_080
        988.0
        0.1
        A_thaliana_081
        3832
        0.3
        A_thaliana_082
        3326
        0.3
        A_thaliana_083
        6129
        0.5
        A_thaliana_084
        7869
        0.7
        A_thaliana_085
        10282
        0.9
        C_hirsuta_086
        2128
        0.2
        C_hirsuta_087
        2760
        0.2
        C_hirsuta_088
        1236
        0.1
        C_hirsuta_089
        1674
        0.1
        C_hirsuta_090
        5036
        0.4
        C_hirsuta_091
        1397
        0.1
        C_hirsuta_092
        4949
        0.4
        C_hirsuta_093
        4653
        0.4
        C_hirsuta_094
        1845
        0.2
        C_hirsuta_095
        2522
        0.2
        C_hirsuta_096
        3759
        0.3
        C_hirsuta_097
        4780
        0.4
        C_hirsuta_098
        1551
        0.1
        C_hirsuta_099
        1823
        0.2
        C_hirsuta_100
        1779
        0.2
        C_hirsuta_101
        2812
        0.2
        C_hirsuta_102
        4061
        0.3
        C_hirsuta_103
        2321
        0.2
        C_hirsuta_104
        3674
        0.3
        C_hirsuta_105
        2202
        0.2
        C_hirsuta_106
        7732
        0.7
        C_hirsuta_107
        1371
        0.1
        C_hirsuta_108
        4480
        0.4
        C_hirsuta_109
        2308
        0.2
        C_hirsuta_110
        4894
        0.4
        C_hirsuta_111
        3548
        0.3
        C_hirsuta_112
        1814
        0.2
        C_hirsuta_113
        2347
        0.2
        C_hirsuta_114
        7081
        0.6
        C_hirsuta_115
        1195
        0.1
        C_hirsuta_116
        3033
        0.3
        C_hirsuta_117
        2102
        0.2
        C_hirsuta_118
        1957
        0.2
        C_hirsuta_119
        3165
        0.3
        C_hirsuta_120
        2427
        0.2
        C_hirsuta_121
        3665
        0.3
        C_hirsuta_122
        1170
        0.1
        C_hirsuta_123
        2309
        0.2
        C_hirsuta_124
        2456
        0.2
        C_hirsuta_125
        2103
        0.2
        C_hirsuta_126
        7490
        0.6
        C_hirsuta_127
        9419
        0.8
        C_hirsuta_128
        3821
        0.3
        C_hirsuta_129
        3506
        0.3
        C_hirsuta_130
        4804
        0.4
        C_hirsuta_131
        4186
        0.4
        C_hirsuta_132
        3728
        0.3
        C_hirsuta_133
        3879
        0.3
        C_hirsuta_134
        2945
        0.2
        C_hirsuta_135
        3734
        0.3
        C_hirsuta_136
        4478
        0.4
        C_hirsuta_137
        2988
        0.3
        C_hirsuta_138
        6051
        0.5
        C_hirsuta_139
        1356
        0.1
        C_hirsuta_140
        2223
        0.2
        C_hirsuta_141
        1769
        0.1
        C_hirsuta_142
        658.0
        0.1
        C_hirsuta_143
        3879
        0.3
        C_hirsuta_144
        5542
        0.5
        C_hirsuta_145
        2067
        0.2
        C_hirsuta_146
        6803
        0.6
        C_hirsuta_147
        2192
        0.2
        C_hirsuta_148
        3711
        0.3
        C_hirsuta_149
        1528
        0.1
        C_hirsuta_150
        8830
        0.8
        C_hirsuta_151
        2576
        0.2
        C_hirsuta_152
        3181
        0.3
        C_hirsuta_153
        4514
        0.4
        C_hirsuta_154
        7527
        0.6
        C_hirsuta_155
        5268
        0.4
        C_hirsuta_156
        6583
        0.6
        C_hirsuta_157
        3132
        0.3
        C_hirsuta_158
        3980
        0.3
        C_hirsuta_159
        5936
        0.5
        C_hirsuta_160
        8381
        0.7
        C_hirsuta_161
        2848
        0.2
        C_hirsuta_162
        4484
        0.4
        C_hirsuta_163
        6504
        0.6
        C_hirsuta_164
        7205
        0.6
        C_hirsuta_165
        6004
        0.5
        C_hirsuta_166
        5245
        0.4
        C_hirsuta_167
        2756
        0.2
        C_hirsuta_168
        876.0
        0.1
        C_hirsuta_169
        6661
        0.6
        C_hirsuta_170
        4453
        0.4
        C_hirsuta_171
        3873
        0.3
        C_hirsuta_172
        2341
        0.2
        D_verna_173
        1787
        0.2
        D_verna_174
        2178
        0.2
        D_verna_175
        2168
        0.2
        D_verna_176
        2620
        0.2
        D_verna_177
        1779
        0.2
        D_verna_178
        928.0
        0.1
        D_verna_180
        1997
        0.2
        D_verna_181
        924.0
        0.1
        D_verna_182
        3074
        0.3
        D_verna_183
        2141
        0.2
        D_verna_184
        2117
        0.2
        D_verna_185
        11817
        1.0
        D_verna_186
        2006
        0.2
        D_verna_187
        3193
        0.3
        D_verna_188
        2472
        0.2
        D_verna_189
        2137
        0.2
        D_verna_190
        3209
        0.3
        D_verna_191
        3336
        0.3
        D_verna_192
        3594
        0.3
        D_verna_193
        1982
        0.2
        D_verna_194
        2850
        0.2
        D_verna_195
        3564
        0.3
        D_verna_196
        3115
        0.3
        D_verna_198
        2917
        0.2
        D_verna_199
        1941
        0.2
        D_verna_200
        2450
        0.2
        D_verna_201
        1935
        0.2
        D_verna_202
        1500
        0.1
        D_verna_203
        1646
        0.1
        D_verna_204
        1225
        0.1
        D_verna_205
        1076
        0.1
        D_verna_206
        2415
        0.2
        D_verna_207
        2282
        0.2
        D_verna_208
        1750
        0.1
        D_verna_209
        1904
        0.2
        D_verna_210
        2085
        0.2
        D_verna_211
        2011
        0.2
        D_verna_212
        1827
        0.2
        D_verna_213
        4715
        0.4
        D_verna_214
        4334
        0.4
        D_verna_215
        5119
        0.4
        D_verna_216
        6872
        0.6
        D_verna_217
        6231
        0.5
        D_verna_218
        6249
        0.5
        D_verna_219
        3267
        0.3
        D_verna_220
        879.0
        0.1
        D_verna_221
        1092
        0.1
        D_verna_222
        6081
        0.5
        D_verna_223
        780.0
        0.1
        D_verna_224
        5409
        0.5
        D_verna_225
        1048
        0.1
        D_verna_226
        1028
        0.1
        D_verna_227
        2985
        0.3
        D_verna_228
        4581
        0.4
        D_verna_229
        3968
        0.3
        D_verna_230
        4334
        0.4
        D_verna_231
        2503
        0.2
        D_verna_232
        3149
        0.3
        D_verna_233
        4991
        0.4
        D_verna_234
        222.0
        0.0
        D_verna_235
        8316
        0.7
        D_verna_236
        7739
        0.7
        D_verna_237
        7265
        0.6
        D_verna_238
        4572
        0.4
        D_verna_239
        8379
        0.7
        D_verna_240
        6495
        0.6
        D_verna_241
        8582
        0.7
        D_verna_242
        10241
        0.9
        D_verna_243
        1307
        0.1
        D_verna_244
        10298
        0.9
        D_verna_245
        10218
        0.9
        D_verna_246
        10914
        0.9
        D_verna_247
        7762
        0.7
        D_verna_248
        3346
        0.3
        D_verna_249
        6451
        0.5
        D_verna_250
        2932
        0.2
        D_verna_251
        7293
        0.6
        D_verna_253
        8156
        0.7
        D_verna_254
        3581
        0.3
        D_verna_255
        0.0
        0.0
        D_verna_256
        6423
        0.5
        D_verna_257
        6608
        0.6
        D_verna_258
        6656
        0.6
        D_verna_259
        7177
        0.6
        D_verna_260
        6629
        0.6
        D_verna_261
        3530
        0.3

        Barcodes of Undetermined Reads


        We have determined the barcodes of your undetermined reads. Here are the top 20 barcodes. The full list is available here. If your libraries are dual indexed, the two indices are concatenated.

        Showing 20/20 rows and 2/2 columns.
        Barcode Sequence(s)CountFrequency (%)
        AACGCTGAATATACAC
        7543.0
        3.1
        CGTAGCGAGCTCTAGT
        6156.0
        2.5
        AACGCTGAACGACGTG
        5295.0
        2.1
        CGTAGCGATGCGTACG
        4537.0
        1.8
        CGAAGTATGATCGTGT
        3732.0
        1.5
        AACGCTGACGTCGCTA
        3717.0
        1.5
        CGTAGCGAGACACTGA
        3432.0
        1.4
        GCGTATACATCGTACG
        3239.0
        1.3
        CGTAGCGATAGTGTAG
        3096.0
        1.3
        CGAAGTATGTCAGATA
        2933.0
        1.2
        CGAAGTATTCGACGAG
        2075.0
        0.8
        TGCTCGTACTACTATA
        2017.0
        0.8
        CGAAGTATACGTCTCG
        1715.0
        0.7
        TGCTCGTACGTTACTA
        1676.0
        0.7
        AGCATACCTCATCGAG
        1275.0
        0.5
        ACTCACTGTCTTTCCC
        1262.0
        0.5
        TGCTCGTATACGAGAC
        1042.0
        0.4
        TGCTCGTAAGAGTCAC
        1039.0
        0.4
        TCTAGACTTCTTTCCC
        946.0
        0.4
        ACGCTACTTCTTTCCC
        932.0
        0.4

        Run Statistics

        Showing 1/1 rows and 4/4 columns.
        LaneTotal # of Single-End ReadsTotal # PF Reads% Undetermined% PhiX Aligned
        1.0
        1286374
        1177607
        20.9
        11.0

        FastQC

        FastQC is a quality control tool for high throughput sequence data, written by Simon Andrews at the Babraham Institute in Cambridge.

        Sequence Counts

        Sequence counts for each sample. Duplicate read counts are an estimate only.

        This plot show the total number of reads, broken down into unique and duplicate if possible (only more recent versions of FastQC give duplicate info).

        You can read more about duplicate calculation in the FastQC documentation. A small part has been copied here for convenience:

        Only sequences which first appear in the first 100,000 sequences in each file are analysed. This should be enough to get a good impression for the duplication levels in the whole file. Each sequence is tracked to the end of the file to give a representative count of the overall duplication level.

        The duplication detection requires an exact sequence match over the whole length of the sequence. Any reads over 75bp in length are truncated to 50bp for this analysis.

        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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