<html>
  <head>
    <meta content="text/html; charset=utf-8" http-equiv="Content-Type">
  </head>
  <body bgcolor="#FFFFFF" text="#000000">
    <p>
      <meta http-equiv="Content-Type" content="text/html; charset=utf-8">
    </p>
    <p>
      <meta name="ProgId" content="Word.Document">
      <meta name="Generator" content="Microsoft Word 12">
      <meta name="Originator" content="Microsoft Word 12">
      <link rel="File-List"
href="file:///C:%5CUsers%5Camine%5CAppData%5CLocal%5CTemp%5Cmsohtmlclip1%5C01%5Cclip_filelist.xml">
      <link rel="themeData"
href="file:///C:%5CUsers%5Camine%5CAppData%5CLocal%5CTemp%5Cmsohtmlclip1%5C01%5Cclip_themedata.thmx">
      <link rel="colorSchemeMapping"
href="file:///C:%5CUsers%5Camine%5CAppData%5CLocal%5CTemp%5Cmsohtmlclip1%5C01%5Cclip_colorschememapping.xml">
      <!--[if gte mso 9]><xml>
 <w:WordDocument>
  <w:View>Normal</w:View>
  <w:Zoom>0</w:Zoom>
  <w:TrackMoves/>
  <w:TrackFormatting/>
  <w:HyphenationZone>21</w:HyphenationZone>
  <w:PunctuationKerning/>
  <w:ValidateAgainstSchemas/>
  <w:SaveIfXMLInvalid>false</w:SaveIfXMLInvalid>
  <w:IgnoreMixedContent>false</w:IgnoreMixedContent>
  <w:AlwaysShowPlaceholderText>false</w:AlwaysShowPlaceholderText>
  <w:DoNotPromoteQF/>
  <w:LidThemeOther>FR</w:LidThemeOther>
  <w:LidThemeAsian>X-NONE</w:LidThemeAsian>
  <w:LidThemeComplexScript>X-NONE</w:LidThemeComplexScript>
  <w:Compatibility>
   <w:BreakWrappedTables/>
   <w:SnapToGridInCell/>
   <w:WrapTextWithPunct/>
   <w:UseAsianBreakRules/>
   <w:DontGrowAutofit/>
   <w:SplitPgBreakAndParaMark/>
   <w:DontVertAlignCellWithSp/>
   <w:DontBreakConstrainedForcedTables/>
   <w:DontVertAlignInTxbx/>
   <w:Word11KerningPairs/>
   <w:CachedColBalance/>
  </w:Compatibility>
  <w:BrowserLevel>MicrosoftInternetExplorer4</w:BrowserLevel>
  <m:mathPr>
   <m:mathFont m:val="Cambria Math"/>
   <m:brkBin m:val="before"/>
   <m:brkBinSub m:val="--"/>
   <m:smallFrac m:val="off"/>
   <m:dispDef/>
   <m:lMargin m:val="0"/>
   <m:rMargin m:val="0"/>
   <m:defJc m:val="centerGroup"/>
   <m:wrapIndent m:val="1440"/>
   <m:intLim m:val="subSup"/>
   <m:naryLim m:val="undOvr"/>
  </m:mathPr></w:WordDocument>
</xml><![endif]--><!--[if gte mso 9]><xml>
 <w:LatentStyles DefLockedState="false" DefUnhideWhenUsed="true"
  DefSemiHidden="true" DefQFormat="false" DefPriority="99"
  LatentStyleCount="267">
  <w:LsdException Locked="false" Priority="0" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Normal"/>
  <w:LsdException Locked="false" Priority="9" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="heading 1"/>
  <w:LsdException Locked="false" Priority="9" QFormat="true" Name="heading 2"/>
  <w:LsdException Locked="false" Priority="9" QFormat="true" Name="heading 3"/>
  <w:LsdException Locked="false" Priority="9" QFormat="true" Name="heading 4"/>
  <w:LsdException Locked="false" Priority="9" QFormat="true" Name="heading 5"/>
  <w:LsdException Locked="false" Priority="9" QFormat="true" Name="heading 6"/>
  <w:LsdException Locked="false" Priority="9" QFormat="true" Name="heading 7"/>
  <w:LsdException Locked="false" Priority="9" QFormat="true" Name="heading 8"/>
  <w:LsdException Locked="false" Priority="9" QFormat="true" Name="heading 9"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 1"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 2"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 3"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 4"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 5"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 6"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 7"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 8"/>
  <w:LsdException Locked="false" Priority="39" Name="toc 9"/>
  <w:LsdException Locked="false" Priority="35" QFormat="true" Name="caption"/>
  <w:LsdException Locked="false" Priority="10" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Title"/>
  <w:LsdException Locked="false" Priority="1" Name="Default Paragraph Font"/>
  <w:LsdException Locked="false" Priority="11" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Subtitle"/>
  <w:LsdException Locked="false" Priority="22" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Strong"/>
  <w:LsdException Locked="false" Priority="20" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Emphasis"/>
  <w:LsdException Locked="false" Priority="59" SemiHidden="false"
   UnhideWhenUsed="false" Name="Table Grid"/>
  <w:LsdException Locked="false" UnhideWhenUsed="false" Name="Placeholder Text"/>
  <w:LsdException Locked="false" Priority="1" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="No Spacing"/>
  <w:LsdException Locked="false" Priority="60" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Shading"/>
  <w:LsdException Locked="false" Priority="61" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light List"/>
  <w:LsdException Locked="false" Priority="62" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Grid"/>
  <w:LsdException Locked="false" Priority="63" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 1"/>
  <w:LsdException Locked="false" Priority="64" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 2"/>
  <w:LsdException Locked="false" Priority="65" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 1"/>
  <w:LsdException Locked="false" Priority="66" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 2"/>
  <w:LsdException Locked="false" Priority="67" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 1"/>
  <w:LsdException Locked="false" Priority="68" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 2"/>
  <w:LsdException Locked="false" Priority="69" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 3"/>
  <w:LsdException Locked="false" Priority="70" SemiHidden="false"
   UnhideWhenUsed="false" Name="Dark List"/>
  <w:LsdException Locked="false" Priority="71" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Shading"/>
  <w:LsdException Locked="false" Priority="72" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful List"/>
  <w:LsdException Locked="false" Priority="73" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Grid"/>
  <w:LsdException Locked="false" Priority="60" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Shading Accent 1"/>
  <w:LsdException Locked="false" Priority="61" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light List Accent 1"/>
  <w:LsdException Locked="false" Priority="62" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Grid Accent 1"/>
  <w:LsdException Locked="false" Priority="63" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 1 Accent 1"/>
  <w:LsdException Locked="false" Priority="64" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 2 Accent 1"/>
  <w:LsdException Locked="false" Priority="65" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 1 Accent 1"/>
  <w:LsdException Locked="false" UnhideWhenUsed="false" Name="Revision"/>
  <w:LsdException Locked="false" Priority="34" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="List Paragraph"/>
  <w:LsdException Locked="false" Priority="29" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Quote"/>
  <w:LsdException Locked="false" Priority="30" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Intense Quote"/>
  <w:LsdException Locked="false" Priority="66" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 2 Accent 1"/>
  <w:LsdException Locked="false" Priority="67" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 1 Accent 1"/>
  <w:LsdException Locked="false" Priority="68" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 2 Accent 1"/>
  <w:LsdException Locked="false" Priority="69" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 3 Accent 1"/>
  <w:LsdException Locked="false" Priority="70" SemiHidden="false"
   UnhideWhenUsed="false" Name="Dark List Accent 1"/>
  <w:LsdException Locked="false" Priority="71" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Shading Accent 1"/>
  <w:LsdException Locked="false" Priority="72" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful List Accent 1"/>
  <w:LsdException Locked="false" Priority="73" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Grid Accent 1"/>
  <w:LsdException Locked="false" Priority="60" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Shading Accent 2"/>
  <w:LsdException Locked="false" Priority="61" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light List Accent 2"/>
  <w:LsdException Locked="false" Priority="62" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Grid Accent 2"/>
  <w:LsdException Locked="false" Priority="63" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 1 Accent 2"/>
  <w:LsdException Locked="false" Priority="64" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 2 Accent 2"/>
  <w:LsdException Locked="false" Priority="65" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 1 Accent 2"/>
  <w:LsdException Locked="false" Priority="66" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 2 Accent 2"/>
  <w:LsdException Locked="false" Priority="67" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 1 Accent 2"/>
  <w:LsdException Locked="false" Priority="68" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 2 Accent 2"/>
  <w:LsdException Locked="false" Priority="69" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 3 Accent 2"/>
  <w:LsdException Locked="false" Priority="70" SemiHidden="false"
   UnhideWhenUsed="false" Name="Dark List Accent 2"/>
  <w:LsdException Locked="false" Priority="71" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Shading Accent 2"/>
  <w:LsdException Locked="false" Priority="72" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful List Accent 2"/>
  <w:LsdException Locked="false" Priority="73" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Grid Accent 2"/>
  <w:LsdException Locked="false" Priority="60" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Shading Accent 3"/>
  <w:LsdException Locked="false" Priority="61" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light List Accent 3"/>
  <w:LsdException Locked="false" Priority="62" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Grid Accent 3"/>
  <w:LsdException Locked="false" Priority="63" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 1 Accent 3"/>
  <w:LsdException Locked="false" Priority="64" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 2 Accent 3"/>
  <w:LsdException Locked="false" Priority="65" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 1 Accent 3"/>
  <w:LsdException Locked="false" Priority="66" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 2 Accent 3"/>
  <w:LsdException Locked="false" Priority="67" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 1 Accent 3"/>
  <w:LsdException Locked="false" Priority="68" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 2 Accent 3"/>
  <w:LsdException Locked="false" Priority="69" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 3 Accent 3"/>
  <w:LsdException Locked="false" Priority="70" SemiHidden="false"
   UnhideWhenUsed="false" Name="Dark List Accent 3"/>
  <w:LsdException Locked="false" Priority="71" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Shading Accent 3"/>
  <w:LsdException Locked="false" Priority="72" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful List Accent 3"/>
  <w:LsdException Locked="false" Priority="73" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Grid Accent 3"/>
  <w:LsdException Locked="false" Priority="60" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Shading Accent 4"/>
  <w:LsdException Locked="false" Priority="61" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light List Accent 4"/>
  <w:LsdException Locked="false" Priority="62" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Grid Accent 4"/>
  <w:LsdException Locked="false" Priority="63" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 1 Accent 4"/>
  <w:LsdException Locked="false" Priority="64" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 2 Accent 4"/>
  <w:LsdException Locked="false" Priority="65" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 1 Accent 4"/>
  <w:LsdException Locked="false" Priority="66" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 2 Accent 4"/>
  <w:LsdException Locked="false" Priority="67" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 1 Accent 4"/>
  <w:LsdException Locked="false" Priority="68" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 2 Accent 4"/>
  <w:LsdException Locked="false" Priority="69" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 3 Accent 4"/>
  <w:LsdException Locked="false" Priority="70" SemiHidden="false"
   UnhideWhenUsed="false" Name="Dark List Accent 4"/>
  <w:LsdException Locked="false" Priority="71" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Shading Accent 4"/>
  <w:LsdException Locked="false" Priority="72" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful List Accent 4"/>
  <w:LsdException Locked="false" Priority="73" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Grid Accent 4"/>
  <w:LsdException Locked="false" Priority="60" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Shading Accent 5"/>
  <w:LsdException Locked="false" Priority="61" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light List Accent 5"/>
  <w:LsdException Locked="false" Priority="62" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Grid Accent 5"/>
  <w:LsdException Locked="false" Priority="63" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 1 Accent 5"/>
  <w:LsdException Locked="false" Priority="64" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 2 Accent 5"/>
  <w:LsdException Locked="false" Priority="65" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 1 Accent 5"/>
  <w:LsdException Locked="false" Priority="66" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 2 Accent 5"/>
  <w:LsdException Locked="false" Priority="67" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 1 Accent 5"/>
  <w:LsdException Locked="false" Priority="68" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 2 Accent 5"/>
  <w:LsdException Locked="false" Priority="69" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 3 Accent 5"/>
  <w:LsdException Locked="false" Priority="70" SemiHidden="false"
   UnhideWhenUsed="false" Name="Dark List Accent 5"/>
  <w:LsdException Locked="false" Priority="71" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Shading Accent 5"/>
  <w:LsdException Locked="false" Priority="72" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful List Accent 5"/>
  <w:LsdException Locked="false" Priority="73" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Grid Accent 5"/>
  <w:LsdException Locked="false" Priority="60" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Shading Accent 6"/>
  <w:LsdException Locked="false" Priority="61" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light List Accent 6"/>
  <w:LsdException Locked="false" Priority="62" SemiHidden="false"
   UnhideWhenUsed="false" Name="Light Grid Accent 6"/>
  <w:LsdException Locked="false" Priority="63" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 1 Accent 6"/>
  <w:LsdException Locked="false" Priority="64" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Shading 2 Accent 6"/>
  <w:LsdException Locked="false" Priority="65" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 1 Accent 6"/>
  <w:LsdException Locked="false" Priority="66" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium List 2 Accent 6"/>
  <w:LsdException Locked="false" Priority="67" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 1 Accent 6"/>
  <w:LsdException Locked="false" Priority="68" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 2 Accent 6"/>
  <w:LsdException Locked="false" Priority="69" SemiHidden="false"
   UnhideWhenUsed="false" Name="Medium Grid 3 Accent 6"/>
  <w:LsdException Locked="false" Priority="70" SemiHidden="false"
   UnhideWhenUsed="false" Name="Dark List Accent 6"/>
  <w:LsdException Locked="false" Priority="71" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Shading Accent 6"/>
  <w:LsdException Locked="false" Priority="72" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful List Accent 6"/>
  <w:LsdException Locked="false" Priority="73" SemiHidden="false"
   UnhideWhenUsed="false" Name="Colorful Grid Accent 6"/>
  <w:LsdException Locked="false" Priority="19" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Subtle Emphasis"/>
  <w:LsdException Locked="false" Priority="21" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Intense Emphasis"/>
  <w:LsdException Locked="false" Priority="31" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Subtle Reference"/>
  <w:LsdException Locked="false" Priority="32" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Intense Reference"/>
  <w:LsdException Locked="false" Priority="33" SemiHidden="false"
   UnhideWhenUsed="false" QFormat="true" Name="Book Title"/>
  <w:LsdException Locked="false" Priority="37" Name="Bibliography"/>
  <w:LsdException Locked="false" Priority="39" QFormat="true" Name="TOC Heading"/>
 </w:LatentStyles>
</xml><![endif]-->
      <style>
<!--
 /* Font Definitions */
 @font-face
        {font-family:"Cambria Math";
        panose-1:2 4 5 3 5 4 6 3 2 4;
        mso-font-charset:0;
        mso-generic-font-family:roman;
        mso-font-pitch:variable;
        mso-font-signature:-536870145 1107305727 0 0 415 0;}
@font-face
        {font-family:Calibri;
        panose-1:2 15 5 2 2 2 4 3 2 4;
        mso-font-charset:0;
        mso-generic-font-family:swiss;
        mso-font-pitch:variable;
        mso-font-signature:-520092929 1073786111 9 0 415 0;}
 /* Style Definitions */
 p.MsoNormal, li.MsoNormal, div.MsoNormal
        {mso-style-unhide:no;
        mso-style-qformat:yes;
        mso-style-parent:"";
        margin-top:0cm;
        margin-right:0cm;
        margin-bottom:10.0pt;
        margin-left:0cm;
        line-height:115%;
        mso-pagination:widow-orphan;
        font-size:11.0pt;
        font-family:"Calibri","sans-serif";
        mso-ascii-font-family:Calibri;
        mso-ascii-theme-font:minor-latin;
        mso-fareast-font-family:Calibri;
        mso-fareast-theme-font:minor-latin;
        mso-hansi-font-family:Calibri;
        mso-hansi-theme-font:minor-latin;
        mso-bidi-font-family:"Times New Roman";
        mso-bidi-theme-font:minor-bidi;
        mso-fareast-language:EN-US;}
.MsoChpDefault
        {mso-style-type:export-only;
        mso-default-props:yes;
        mso-ascii-font-family:Calibri;
        mso-ascii-theme-font:minor-latin;
        mso-fareast-font-family:Calibri;
        mso-fareast-theme-font:minor-latin;
        mso-hansi-font-family:Calibri;
        mso-hansi-theme-font:minor-latin;
        mso-bidi-font-family:"Times New Roman";
        mso-bidi-theme-font:minor-bidi;
        mso-fareast-language:EN-US;}
.MsoPapDefault
        {mso-style-type:export-only;
        margin-bottom:10.0pt;
        line-height:115%;}
@page Section1
        {size:612.0pt 792.0pt;
        margin:70.85pt 70.85pt 70.85pt 70.85pt;
        mso-header-margin:36.0pt;
        mso-footer-margin:36.0pt;
        mso-paper-source:0;}
div.Section1
        {page:Section1;}
 /* List Definitions */
 @list l0
        {mso-list-id:1874493137;
        mso-list-template-ids:1610103466;}
@list l0:level1
        {mso-level-number-format:bullet;
        mso-level-text:;
        mso-level-tab-stop:36.0pt;
        mso-level-number-position:left;
        text-indent:-18.0pt;
        mso-ansi-font-size:10.0pt;
        font-family:Symbol;}
ol
        {margin-bottom:0cm;}
ul
        {margin-bottom:0cm;}
-->
</style><!--[if gte mso 10]>
<style>
 /* Style Definitions */
 table.MsoNormalTable
        {mso-style-name:"Tableau Normal";
        mso-tstyle-rowband-size:0;
        mso-tstyle-colband-size:0;
        mso-style-noshow:yes;
        mso-style-priority:99;
        mso-style-qformat:yes;
        mso-style-parent:"";
        mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
        mso-para-margin-top:0cm;
        mso-para-margin-right:0cm;
        mso-para-margin-bottom:10.0pt;
        mso-para-margin-left:0cm;
        line-height:115%;
        mso-pagination:widow-orphan;
        font-size:11.0pt;
        font-family:"Calibri","sans-serif";
        mso-ascii-font-family:Calibri;
        mso-ascii-theme-font:minor-latin;
        mso-hansi-font-family:Calibri;
        mso-hansi-theme-font:minor-latin;
        mso-bidi-font-family:"Times New Roman";
        mso-bidi-theme-font:minor-bidi;
        mso-fareast-language:EN-US;}
</style>
<![endif]-->
      <p class="MsoNormal"><span
          style="font-size:12.0pt;line-height:115%;
          font-family:"Times New
          Roman","serif";mso-ansi-language:EN-US"
          lang="EN-US">Thank you <span
            style="color:black;background:white">Michael <span
              style="mso-spacerun:yes"> </span>for your response. <o:p></o:p></span></span></p>
      <p class="MsoNormal"><span
          style="font-size:12.0pt;line-height:115%;
          font-family:"Times New
Roman","serif";color:black;background:white;mso-ansi-language:EN-US"
          lang="EN-US">As suggested by you, I would use Augustus and<span
            style="mso-spacerun:yes">  </span>Snap trained both by the
          assembled
          transcripts in a bootstrap fashion.<o:p></o:p></span></p>
      <p class="MsoNormal"><span
          style="font-size:12.0pt;line-height:115%;
          font-family:"Times New
Roman","serif";color:black;background:white;mso-ansi-language:EN-US"
          lang="EN-US">For the masking, I intend to to adapt <span
            style="mso-spacerun:yes"> </span>Carson</span> strategy :<o:p></o:p></p>
      <p class="MsoNormal"
        style="mso-margin-top-alt:auto;margin-bottom:1.2pt;
margin-left:19.2pt;text-indent:-18.0pt;line-height:18.0pt;mso-list:l0
        level1 lfo1;
        tab-stops:list 36.0pt;background:white"><!--[if !supportLists]--><span
style="font-size:10.0pt;mso-bidi-font-size:12.0pt;font-family:Symbol;
mso-fareast-font-family:Symbol;mso-bidi-font-family:Symbol;color:black;
          mso-ansi-language:EN-US;mso-fareast-language:FR" lang="EN-US"><span
            style="mso-list:Ignore">·<span style="font:7.0pt "Times
              New Roman"">        
            </span></span></span><!--[endif]--><span
          style="font-size:12.0pt;
          font-family:"Times New
          Roman","serif";mso-fareast-font-family:"Times
          New Roman";
          color:black;mso-ansi-language:EN-US;mso-fareast-language:FR"
          lang="EN-US">Collecting
          RepeatModeler repeats.lib<o:p></o:p></span></p>
      <p class="MsoNormal"
        style="mso-margin-top-alt:auto;margin-bottom:1.2pt;
margin-left:19.2pt;text-indent:-18.0pt;line-height:18.0pt;mso-list:l0
        level1 lfo1;
        tab-stops:list 36.0pt;background:white"><!--[if !supportLists]--><span
style="font-size:10.0pt;mso-bidi-font-size:12.0pt;font-family:Symbol;
mso-fareast-font-family:Symbol;mso-bidi-font-family:Symbol;mso-ansi-language:
          EN-US" lang="EN-US"><span style="mso-list:Ignore">·<span
              style="font:7.0pt "Times New Roman"">        
            </span></span></span><!--[endif]--><span
          style="font-size:12.0pt;
          font-family:"Times New
          Roman","serif";mso-fareast-font-family:"Times
          New Roman";
          mso-ansi-language:EN-US;mso-fareast-language:FR" lang="EN-US">Searching
          Sequences in
          Modelerunknown.lib <span style="mso-spacerun:yes"> </span>against
          a transposase
          database (derived from </span><span
          style="font-size:12.0pt;font-family:
          "Times New
          Roman","serif";mso-fareast-font-family:"Times
          New Roman";mso-fareast-language:
          FR"><a href="http://www.repeatmasker.org/"><span
              style="color:windowtext;
mso-ansi-language:EN-US;text-decoration:none;text-underline:none"
              lang="EN-US">RepeatMasker</span></a></span><span
          style="font-size:12.0pt;font-family:"Times New
          Roman","serif";
          mso-fareast-font-family:"Times New
          Roman";mso-ansi-language:EN-US;mso-fareast-language:
          FR" lang="EN-US"> package and </span><span
          style="font-size:12.0pt;font-family:
          "Times New
          Roman","serif";mso-fareast-font-family:"Times
          New Roman";mso-fareast-language:
          FR"><a href="http://www.ncbi.nlm.nih.gov/pubmed/21535899"><span
style="color:windowtext;mso-ansi-language:EN-US;text-decoration:none;
              text-underline:none" lang="EN-US">Kennedy et al (2011)</span></a></span><span
          style="font-size:12.0pt;font-family:"Times New
          Roman","serif";mso-fareast-font-family:
          "Times New
          Roman";mso-ansi-language:EN-US;mso-fareast-language:FR"
          lang="EN-US">) and <span style="mso-spacerun:yes"> </span>considering
          sequences matching <span style="mso-spacerun:yes"> </span>transposases
          as transposons.</span><span
          style="font-size:12.0pt;font-family:"Times New
          Roman","serif";
          mso-ansi-language:EN-US" lang="EN-US"><o:p></o:p></span></p>
      <p class="MsoNormal"
        style="mso-margin-top-alt:auto;margin-bottom:1.2pt;
margin-left:19.2pt;text-indent:-18.0pt;line-height:18.0pt;mso-list:l0
        level1 lfo1;
        tab-stops:list 36.0pt;background:white"><!--[if !supportLists]--><span
style="font-size:10.0pt;mso-bidi-font-size:12.0pt;font-family:Symbol;
mso-fareast-font-family:Symbol;mso-bidi-font-family:Symbol;mso-ansi-language:
          EN-US" lang="EN-US"><span style="mso-list:Ignore">·<span
              style="font:7.0pt "Times New Roman"">        
            </span></span></span><!--[endif]--><span
          style="font-size:12.0pt;
          font-family:"Times New
          Roman","serif";mso-ansi-language:EN-US"
          lang="EN-US">Exclusion of
          gene fragments in both known and unkown repeats<o:p></o:p></span></p>
      <p class="MsoNormal"
        style="mso-margin-top-alt:auto;margin-bottom:1.2pt;
margin-left:19.2pt;text-indent:-18.0pt;line-height:18.0pt;mso-list:l0
        level1 lfo1;
        tab-stops:list 36.0pt;background:white"><!--[if !supportLists]--><span
style="font-size:10.0pt;mso-bidi-font-size:12.0pt;font-family:Symbol;
mso-fareast-font-family:Symbol;mso-bidi-font-family:Symbol;mso-ansi-language:
          EN-US" lang="EN-US"><span style="mso-list:Ignore">·<span
              style="font:7.0pt "Times New Roman"">        
            </span></span></span><!--[endif]--><span
          style="font-size:12.0pt;
          font-family:"Times New
          Roman","serif";mso-ansi-language:EN-US"
          lang="EN-US">As I'm concerned
          by gene duplications, the remainder sequences in the unkown <span
            style="mso-spacerun:yes"> </span>lib present less than 10
          times will be
          removed.<o:p></o:p></span></p>
      <p class="MsoNormal"
        style="mso-margin-top-alt:auto;margin-bottom:1.2pt;
        margin-left:19.2pt;line-height:18.0pt;background:white"><span
          style="font-size:12.0pt;font-family:"Times New
          Roman","serif";mso-ansi-language:
          EN-US" lang="EN-US"><br>
        </span></p>
      <p class="MsoNormal"
        style="mso-margin-top-alt:auto;margin-bottom:1.2pt;
        margin-left:19.2pt;line-height:18.0pt;background:white"><span
          style="font-size:12.0pt;font-family:"Times New
          Roman","serif";mso-ansi-language:
          EN-US" lang="EN-US">Thank you again for your time and I remain
          open to any suggestion.<o:p></o:p></span></p>
      <p class="MsoNormal"
        style="mso-margin-top-alt:auto;margin-bottom:1.2pt;
        margin-left:19.2pt;line-height:18.0pt;background:white"><span
          style="font-size:12.0pt;font-family:"Times New
          Roman","serif";mso-ansi-language:
          EN-US" lang="EN-US"><o:p> </o:p></span></p>
      <p class="MsoNormal"
        style="mso-margin-top-alt:auto;margin-bottom:1.2pt;
        margin-left:19.2pt;line-height:18.0pt;background:white"><span
          style="font-size:12.0pt;font-family:"Times New
          Roman","serif";mso-ansi-language:
          EN-US" lang="EN-US">Best,<o:p></o:p></span></p>
      <span
        style="font-size:12.0pt;line-height:115%;font-family:"Times
        New Roman","serif";
mso-fareast-font-family:Calibri;mso-fareast-theme-font:minor-latin;mso-ansi-language:
        EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA"
        lang="EN-US">Amine</span></p>
    <br>
    <div class="moz-cite-prefix">Le 27/10/2016 à 15:21, Michael Campbell
      a écrit :<br>
    </div>
    <blockquote
      cite="mid:8935E6BD-FDEC-464B-B174-94649CB42D63@gmail.com"
      type="cite">
      <meta http-equiv="Content-Type" content="text/html; charset=utf-8">
      I think that if you train any further you will run the risk of
      overtraining. setting alt_splice to 1 will add transcripts but not
      genes, so the gene count is going to be related to the training of
      the gene finder. I would recommend looking at a few of your large
      scaffolds in a genome browser. I would also recommend adding a
      second gene predictor such as augustus. When multiple predictors
      are used and the models they predict converge you can have more
      confidence in the gene prediction.
      <div class=""><br class="">
      </div>
      <div class="">For the masking you can make a species specific
        repeat library like Carson suggested to see if the gene count
        comes down a little. If you are concerned about masking
        duplicated genes you cad do a couple of things. You can filter
        the repeat library based on known proteins. You can also set a
        copy number minimum for the making and only include repeats that
        are present more than 10 time in the genome. Here are a couple
        of URLs for making species specific repeat libraries </div>
      <div class=""><a moz-do-not-send="true"
href="http://weatherby.genetics.utah.edu/MAKER/wiki/index.php/Repeat_Library_Construction-Advanced"
          class="">http://weatherby.genetics.utah.edu/MAKER/wiki/index.php/Repeat_Library_Construction-Advanced</a></div>
      <div class=""><a moz-do-not-send="true"
href="http://weatherby.genetics.utah.edu/MAKER/wiki/index.php/Repeat_Library_Construction-Basic"
          class="">http://weatherby.genetics.utah.edu/MAKER/wiki/index.php/Repeat_Library_Construction-Basic</a></div>
      <div class=""><br class="">
      </div>
      <div class="">Take care,</div>
      <div class="">Mike</div>
      <div class=""> <br class="">
        <div>
          <blockquote type="cite" class="">
            <div class="">On Oct 27, 2016, at 5:54 AM, Mohamed Amine
              CHEBBI <<a moz-do-not-send="true"
                href="mailto:mohamed.amine.chebbi@univ-poitiers.fr"
                class="">mohamed.amine.chebbi@univ-poitiers.fr</a>>
              wrote:</div>
            <br class="Apple-interchange-newline">
            <div class="">
              <div style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class=""><br
                  class="webkit-block-placeholder">
              </div>
              <div style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class=""><br
                  class="webkit-block-placeholder">
              </div>
              <div style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class=""><br
                  class="webkit-block-placeholder">
              </div>
              <p class="MsoNormal" style="margin: 0cm 0cm 10pt;
                line-height: normal; font-size: 11pt; font-family:
                Calibri, sans-serif; font-style: normal; font-variant:
                normal; font-weight: normal; letter-spacing: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);"><span
                  style="font-size: 14pt;" class="" lang="EN-US">Sorry,
                  the X and Y were switched in the plot due to a
                  mishandling. Please find attached now the correct AED
                  graph.<span class="Apple-converted-space"> </span><o:p
                    class=""></o:p></span></p>
              <p class="MsoNormal" style="margin: 0cm 0cm 10pt;
                line-height: normal; font-size: 11pt; font-family:
                Calibri, sans-serif; font-style: normal; font-variant:
                normal; font-weight: normal; letter-spacing: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);"><span
                  style="font-size: 14pt;" class="" lang="EN-US">The
                  round 3 (red curve) shows little higher overall AED
                  than the second round (green curve) and more genes (<span
                    style="color: rgb(33, 33, 33); background-color:
                    white; background-position: initial initial;
                    background-repeat: initial initial;" class="">22931
                    comparing to 22547 in the round 2). Do you think
                    that I should stop at the second round ?</span><o:p
                    class=""></o:p></span></p>
              <p class="MsoNormal" style="margin: 0cm 0cm 10pt;
                line-height: normal; font-size: 11pt; font-family:
                Calibri, sans-serif; font-style: normal; font-variant:
                normal; font-weight: normal; letter-spacing: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);"><span
                  style="font-size: 14pt;" class="" lang="EN-US">I
                  didn't<span class=""> <span
                      class="Apple-converted-space"> </span></span>precise
                  in the precedent email that the Repeat masking was
                  done in Maker using the Repbase and only models found
                  by RepeatModeler having identities. I let<span
                    class=""> <span class="Apple-converted-space"> </span></span>unmasked
                  the unkown lib of RepeatModeler. In fact we expect a
                  high rate of segmental and gene duplication in the
                  genome and then we  could explain the high overall
                  count of genes found by Maker.<o:p class=""></o:p></span></p>
              <p class="MsoNormal" style="margin: 0cm 0cm 10pt;
                line-height: normal; font-size: 11pt; font-family:
                Calibri, sans-serif; font-style: normal; font-variant:
                normal; font-weight: normal; letter-spacing: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);"><span
                  style="font-size: 14pt;" class="" lang="EN-US">In the
                  other hand the high, rate of genes may be also
                  expalined by the fact that I activate the alt_splice=1
                  option to find alternative splicing, do you think that
                  it was a good idea ?<o:p class=""></o:p></span></p>
              <p class="MsoNormal" style="margin: 0cm 0cm 10pt;
                line-height: normal; font-size: 11pt; font-family:
                Calibri, sans-serif; font-style: normal; font-variant:
                normal; font-weight: normal; letter-spacing: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);"><span
                  style="font-size: 14pt;" class="" lang="EN-US"> <o:p
                    class=""></o:p></span></p>
              <p class="MsoNormal" style="margin: 0cm 0cm 10pt;
                line-height: normal; font-size: 11pt; font-family:
                Calibri, sans-serif; font-style: normal; font-variant:
                normal; font-weight: normal; letter-spacing: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);"><span
                  style="font-size: 14pt;" class="" lang="EN-US">Thank
                  you very much for your time.<span
                    class="Apple-converted-space"> </span><o:p class=""></o:p></span></p>
              <br style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class="">
              <div style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class=""><br
                  class="webkit-block-placeholder">
              </div>
              <p style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class=""><span
                  class="" lang="EN-US">Best,</span></p>
              <p class="MsoNormal" style="margin: 0cm 0cm 10pt;
                line-height: 16.866666793823242px; font-size: 11pt;
                font-family: Calibri, sans-serif; font-style: normal;
                font-variant: normal; font-weight: normal;
                letter-spacing: normal; orphans: auto; text-align:
                start; text-indent: 0px; text-transform: none;
                white-space: normal; widows: auto; word-spacing: 0px;
                -webkit-text-stroke-width: 0px; background-color:
                rgb(255, 255, 255);">Amine<br class="">
                <span class="" lang="EN-US"><o:p class=""></o:p></span></p>
              <div style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class=""><br
                  class="webkit-block-placeholder">
              </div>
              <br style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class="">
              <div class="moz-cite-prefix" style="font-family:
                Helvetica; font-size: 18px; font-style: normal;
                font-variant: normal; font-weight: normal;
                letter-spacing: normal; line-height: normal; orphans:
                auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);">Le 26/10/2016 à
                20:06, Carson Holt a écrit :<br class="">
              </div>
              <blockquote
                cite="mid:3EA2EC84-9B2A-4631-97F8-44D774E67468@gmail.com"
                type="cite" style="font-family: Helvetica; font-size:
                18px; font-style: normal; font-variant: normal;
                font-weight: normal; letter-spacing: normal;
                line-height: normal; orphans: auto; text-align: start;
                text-indent: 0px; text-transform: none; white-space:
                normal; widows: auto; word-spacing: 0px;
                -webkit-text-stroke-width: 0px; background-color:
                rgb(255, 255, 255);" class="">Sorry. I also assumed X
                and Y was flipped when I looked at it. Now I read the
                labels, your AED curve would be weird unless the X and Y
                are flipped in your figure.
                <div class=""><br class="">
                </div>
                <div class="">—Carson</div>
                <div class=""><br class="">
                  <div class=""><br class="">
                    <div class="">
                      <blockquote type="cite" class="">
                        <div class="">On Oct 26, 2016, at 12:04 PM,
                          Carson Holt <<a moz-do-not-send="true"
                            href="mailto:carsonhh@gmail.com" class="">carsonhh@gmail.com</a>>
                          wrote:</div>
                        <br class="Apple-interchange-newline">
                        <div class="">
                          <div class="" style="word-wrap: break-word;
                            -webkit-nbsp-mode: space;
                            -webkit-line-break: after-white-space;">Your
                            AED curve looks fine. The first run (using
                            protein2genome or est2genome I assume) will
                            always have really low overall AED because
                            they are exact copies of the
                            protein/transcript alignments (so AED is
                            meaningless there because it will always
                            artificially look good). The protein2genome
                            or est2genome modles also have a hard
                            end-to-end coverage filtering cutoff of 0.5
                            when generated (apparent in the curve -
                            value in maker_bopts.ctl). The next runs
                            with SNAP show >80% of models with AED
                            under 0.5, so it looks good. You can further
                            look at models by adding protein domains
                            using InterProScan in which you would expect
                            70-80% of models to contain a recognizable
                            InterPro domain (false and bad models will
                            result in very low overall domain content).
                            <div class=""><br class="">
                            </div>
                            <div class="">Your overall gene counts are a
                              little high though for an arthropod
                              (14,000-19,000 genes would be expected as
                              gene loss rather than gene gain is the
                              primary evolutionary force in the
                              Ecdysozoa). However your gene counts can
                              be explained by either insufficient repeat
                              masking (you can add a RepeatModeler
                              generated library to the existing settings
                              to help with this), poor mRNA-seq assembly
                              or a lot of noise in the RNA-seq (this can
                              be helped with more strict assembly
                              parameters including the jaccard-clip
                              option in trinity), or it is just the
                              result of assembly fragmentation (if you
                              have a lot of contigs or runs of NNNN in
                              the assembly, then many genes will be
                              split which results in inflated gene
                              counts).<br class="">
                              <div class=""><br class="">
                              </div>
                              <div class="">Finally manually look at the
                                most gene dense contigs in a browser
                                like Apollo or IGV (gene_density =
                                gene_count / contig_length). If the most
                                gene dense contigs are overwhelmingly
                                single exon, then you may need to filter
                                out some prokaryotic assembly
                                contamination (not uncommon). If you
                                have contamination, it will assemble as
                                independent contigs, so is easily
                                blacklisted and can be identified
                                visually (always gene dense and single
                                exon).</div>
                              <div class=""><br class="">
                              </div>
                              <div class="">Thanks,</div>
                              <div class="">Carson</div>
                              <div class=""><br class="">
                              </div>
                              <div class=""><br class="">
                              </div>
                              <div class=""><br class="">
                              </div>
                              <div class=""><br class="">
                                <div class="">
                                  <blockquote type="cite" class="">
                                    <div class="">On Oct 26, 2016, at
                                      7:09 AM, Mohamed Amine Chebbi <<a
                                        moz-do-not-send="true"
                                        class="moz-txt-link-abbreviated"
href="mailto:mohamed.amine.chebbi@univ-poitiers.fr"><a class="moz-txt-link-abbreviated" href="mailto:mohamed.amine.chebbi@univ-poitiers.fr">mohamed.amine.chebbi@univ-poitiers.fr</a></a>>
                                      wrote:</div>
                                    <br
                                      class="Apple-interchange-newline">
                                    <div class="">
                                      <div class="">
                                        <div class=""
                                          style="font-family: arial,
                                          helvetica, sans-serif;
                                          font-size: 12pt;">
                                          <div class="">
                                            <div class="" style="margin:
                                              0px; line-height: normal;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222; background:
                                                white;" class=""
                                                style="font-size: 12pt;
                                                color: rgb(34, 34, 34);
                                                background-color: white;
                                                background-position:
                                                initial initial;
                                                background-repeat:
                                                initial initial;">Hi !</span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222;" class=""
                                                style="font-size: 12pt;">I
                                                have tried three rounds
                                                of annotation in Maker
                                                on a non model arthropod
                                                genome (1.7Gb) which is
                                                an hybrid assembly of
                                                Pacbio and illumina
                                                reads.</span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222;" class=""
                                                style="font-size: 12pt;">As
                                                suggested in the
                                                tutorial, I ran in the
                                                first round Maker with
                                                repeat masking to
                                                generate gene models
                                                using transcript
                                                (Trinity assembly) and
                                                protein (swissprot)
                                                evidence. Then Maker
                                                models were used twice
                                                in a bootstrap fashion
                                                to retrain SNAP.  </span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222;" class=""
                                                style="font-size: 12pt;">The
                                                number of genes drops
                                                from<span
                                                  class="Apple-converted-space"> </span></span><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #212121; background:
                                                white;" class=""
                                                style="font-size: 12pt;
                                                color: rgb(33, 33, 33);
                                                background-color: white;
                                                background-position:
                                                initial initial;
                                                background-repeat:
                                                initial initial;">29207
                                                in the round 1 to 22547
                                                in the round 2 then
                                                increases slightly to
                                                22931 in the round 3.</span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><br
                                                class="">
                                              <span
                                                data-mce-style="font-size:
                                                12.0pt; font-family:
                                                'Times New
                                                Roman','serif'; color:
                                                #212121; background:
                                                white;" class=""
                                                style="font-size: 12pt;
                                                color: rgb(33, 33, 33);
                                                background-color: white;
                                                background-position:
                                                initial initial;
                                                background-repeat:
                                                initial initial;"><span data-mce-style="font-variant-ligatures: normal; font-variant-caps: normal; orphans: 2; white-space: pre-wrap; widows: 2; -webkit-text-stroke-width: 0px; float: none; word-spacing: 0px;" class="" style="orphans: 2; white-space: pre-wrap; widows: 2; -webkit-text-stroke-width: 0px; float: none; word-spacing: 0px;">However, the AED profile (attached) don't seem to be satisfactory.</span></span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #212121; background:
                                                white;" class=""
                                                style="font-size: 12pt;
                                                color: rgb(33, 33, 33);
                                                background-color: white;
                                                background-position:
                                                initial initial;
                                                background-repeat:
                                                initial initial;">So I
                                                wonder if you could let
                                                me a good strategy to
                                                improve the annotation
                                                quality. Do you think
                                                that filtering good
                                                transcripts could
                                                improve results. If yes
                                                , which criteria should</span><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222;" class=""
                                                style="font-size: 12pt;"><span
class="Apple-converted-space"> </span>be taken into account ?</span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222;" class=""
                                                style="font-size: 12pt;">Thank
                                                you.</span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222;" class=""
                                                style="font-size: 12pt;"><br
                                                  data-mce-bogus="1"
                                                  class="">
                                              </span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222;" class=""
                                                style="font-size: 12pt;">Best;</span></div>
                                            <div class=""
                                              style="line-height:
                                              normal; background-color:
                                              white; margin: 0px;"><span
data-mce-style="font-size: 12.0pt; font-family: 'Times New
                                                Roman','serif'; color:
                                                #222222;" class=""
                                                style="font-size: 12pt;">Amine</span></div>
                                          </div>
                                        </div>
                                      </div>
                                      <span
                                        id="cid:A35AEA9C-A0B8-490E-84CF-C956A8207A2C@genetics.utah.edu"
                                        class=""><AED-Graph.pdf></span>_______________________________________________<br
                                        class="">
                                      maker-devel mailing list<br
                                        class="">
                                      <a moz-do-not-send="true"
                                        href="mailto:maker-devel@box290.bluehost.com"
                                        class="">maker-devel@box290.bluehost.com</a><br
                                        class="">
                                      <a moz-do-not-send="true"
href="http://box290.bluehost.com/mailman/listinfo/maker-devel_yandell-lab.org"
                                        class="">http://box290.bluehost.com/mailman/listinfo/maker-devel_yandell-lab.org</a><br
                                        class="">
                                    </div>
                                  </blockquote>
                                </div>
                                <br class="">
                              </div>
                            </div>
                          </div>
                        </div>
                      </blockquote>
                    </div>
                    <br class="">
                  </div>
                </div>
              </blockquote>
              <br style="font-family: Helvetica; font-size: 18px;
                font-style: normal; font-variant: normal; font-weight:
                normal; letter-spacing: normal; line-height: normal;
                orphans: auto; text-align: start; text-indent: 0px;
                text-transform: none; white-space: normal; widows: auto;
                word-spacing: 0px; -webkit-text-stroke-width: 0px;
                background-color: rgb(255, 255, 255);" class="">
              <pre class="moz-signature" cols="72" style="font-size: 18px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255);">-- 
Mohamed Amine CHEBBI, PhD Student
Université de Poitiers
Laboratoire Ecologie et Biologie des Interactions - UMR CNRS 7267
Equipe Ecologie Evolution Symbiose
Bât. B8-B35 - 5 Rue Albert Turpin
TSA 51106
F-86022 Poitiers Cedex 9
FRANCE
Lab website: <a moz-do-not-send="true" class="moz-txt-link-freetext" href="http://ecoevol.labo.univ-poitiers.fr/">http://ecoevol.labo.univ-poitiers.fr/</a></pre>
              <span id="cid:A1915C92-1E48-4258-A2BE-A5E560F53C31"><AED-Graph.pdf></span></div>
          </blockquote>
        </div>
        <br class="">
      </div>
    </blockquote>
    <br>
    <pre class="moz-signature" cols="72">-- 
Mohamed Amine CHEBBI, PhD Student
Université de Poitiers
Laboratoire Ecologie et Biologie des Interactions - UMR CNRS 7267
Equipe Ecologie Evolution Symbiose
Bât. B8-B35 - 5 Rue Albert Turpin
TSA 51106
F-86022 Poitiers Cedex 9
FRANCE
Lab website: <a class="moz-txt-link-freetext" href="http://ecoevol.labo.univ-poitiers.fr/">http://ecoevol.labo.univ-poitiers.fr/</a></pre>
  </body>
</html>