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1 <tool id="cshl_fasta_clipping_histogram" name="Length Distribution">
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2 <description>chart</description>
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3 <command>fasta_clipping_histogram.pl $input $outfile</command>
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4
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5 <inputs>
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6 <param format="fasta" name="input" type="data" label="Library to analyze" />
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7 </inputs>
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8
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9 <outputs>
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10 <data format="png" name="outfile" metadata_source="input"
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11 />
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12 </outputs>
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13 <help>
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14
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15 **What it does**
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16
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17 This tool creates a histogram image of sequence lengths distribution in a given fasta dataset file.
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18
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19 **TIP:** Use this tool after clipping your library (with **FASTX Clipper tool**), to visualize the clipping results.
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20
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21 -----
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22
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23 **Output Examples**
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24
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25 In the following library, most sequences are 24-mers to 27-mers.
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26 This could indicate an abundance of endo-siRNAs (depending of course of what you've tried to sequence in the first place).
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27
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28 .. image:: ./static/fastx_icons/fasta_clipping_histogram_1.png
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29
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30
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31 In the following library, most sequences are 19,22 or 23-mers.
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32 This could indicate an abundance of miRNAs (depending of course of what you've tried to sequence in the first place).
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33
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34 .. image:: ./static/fastx_icons/fasta_clipping_histogram_2.png
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35
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36
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37 -----
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38
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39
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40 **Input Formats**
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41
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42 This tool accepts short-reads FASTA files. The reads don't have to be short, but they do have to be on a single line, like so::
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43
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44 >sequence1
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45 AGTAGTAGGTGATGTAGAGAGAGAGAGAGTAG
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46 >sequence2
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47 GTGTGTGTGGGAAGTTGACACAGTA
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48 >sequence3
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49 CCTTGAGATTAACGCTAATCAAGTAAAC
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50
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51
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52 If the sequences span over multiple lines::
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53
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54 >sequence1
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55 CAGCATCTACATAATATGATCGCTATTAAACTTAAATCTCCTTGACGGAG
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56 TCTTCGGTCATAACACAAACCCAGACCTACGTATATGACAAAGCTAATAG
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57 aactggtctttacctTTAAGTTG
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58
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59 Use the **FASTA Width Formatter** tool to re-format the FASTA into a single-lined sequences::
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60
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61 >sequence1
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62 CAGCATCTACATAATATGATCGCTATTAAACTTAAATCTCCTTGACGGAGTCTTCGGTCATAACACAAACCCAGACCTACGTATATGACAAAGCTAATAGaactggtctttacctTTAAGTTG
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63
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64
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65 -----
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66
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67
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68
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69 **Multiplicity counts (a.k.a reads-count)**
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70
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71 If the sequence identifier (the text after the '>') contains a dash and a number, it is treated as a multiplicity count value (i.e. how many times that individual sequence repeated in the original FASTA file, before collapsing).
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72
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73 Example 1 - The following FASTA file *does not* have multiplicity counts::
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74
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75 >seq1
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76 GGATCC
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77 >seq2
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78 GGTCATGGGTTTAAA
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79 >seq3
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80 GGGATATATCCCCACACACACACAC
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81
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82 Each sequence is counts as one, to produce the following chart:
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83
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84 .. image:: ./static/fastx_icons/fasta_clipping_histogram_3.png
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85
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86
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87 Example 2 - The following FASTA file have multiplicity counts::
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88
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89 >seq1-2
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90 GGATCC
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91 >seq2-10
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92 GGTCATGGGTTTAAA
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93 >seq3-3
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94 GGGATATATCCCCACACACACACAC
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95
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96 The first sequence counts as 2, the second as 10, the third as 3, to produce the following chart:
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97
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98 .. image:: ./static/fastx_icons/fasta_clipping_histogram_4.png
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99
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100 Use the **FASTA Collapser** tool to create FASTA files with multiplicity counts.
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101
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102 </help>
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103 </tool>
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104 <!-- FASTA-Clipping-Histogram is part of the FASTX-toolkit, by A.Gordon (gordon@cshl.edu) -->
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