Team:Lethbridge/Notebook

From 2009.igem.org

(Difference between revisions)
(September)
(October)
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<li><a href="https://2009.igem.org/Team:Lethbridge/Criteria" style="width:1px;">Judging</a></li>
<li><a href="https://2009.igem.org/Team:Lethbridge/Criteria" style="width:1px;">Judging</a></li>
<li><a href="https://2009.igem.org/Team:Lethbridge/videos" style="width:125px;">Extra Information on SynBio</a></li>
<li><a href="https://2009.igem.org/Team:Lethbridge/videos" style="width:125px;">Extra Information on SynBio</a></li>
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== October ==
 
-
 
-
===October 1===
 
-
 
-
'''Roxanne'''
 
-
Gel for gel extraction
 
-
 
-
Restriction
 
-
Ribo-Chez-dT
 
-
*2x CheZ-dT
 
-
*2x GFP-dT
 
-
*3x C-EYFP-dT
 
-
 
-
Ligation
 
-
*C-ECFP-dT
 
-
*C-CFP-dT
 
-
*C-EYFP-dT
 
-
*mRBS-N-ECFP
 
-
*mRBS-N-EYFP
 
-
*pSB-C-EFP
 
-
*pSB-C-EYFP
 
-
*pSB-C-CFP
 
-
*pSB-N-ECFP
 
-
*pSB-N-EYFP
 
-
 
-
Aliquoted DH5α cells
 
-
 
-
'''Ashley'''
 
-
 
-
{|
 
-
|Tube
 
-
|'''Tube + Gel'''
 
-
|Gel
 
-
|-
 
-
|1.1226g
 
-
|'''1.4931g'''
 
-
|0.2671g
 
-
|-
 
-
|1.012g
 
-
|'''1.2302g'''
 
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|0.1290g
 
-
|}
 
-
 
-
Eluted with 50uL buffer EB
 
-
 
-
Ligation
 
-
 
-
{|
 
-
|
 
-
|'''1:3'''
 
-
|2:6
 
-
|-
 
-
|lum-dT
 
-
|'''3uL'''
 
-
|6uL
 
-
|-
 
-
|SRBS
 
-
|'''1uL'''
 
-
|2uL
 
-
|-
 
-
|Ligase Buffer
 
-
|'''1uL'''
 
-
|2uL
 
-
|-
 
-
|Water
 
-
|4.5uL
 
-
|9uL
 
-
|-
 
-
|Ligase
 
-
|0.5uL
 
-
|1uL
 
-
|}
 
-
===Oct 2===
 
-
Pick Colonies:
 
-
*sRBS + Lumazine-dT
 
-
*mRBS + N-term FPs
 
-
*C-term FPs + dT
 
-
*pSB + N-term FPs
 
-
*pSB + N-term FPs
 
-
 
-
Gel Extraction:
 
-
*Riboswitch-cheZ-dT
 
-
*2x cheZ-dT (in case Ribo-cheZ-dT isn't actually there)
 
-
*3x C-EYFP-dT
 
-
*2x GFP-dT
 
-
 
-
Liquid Culture:
 
-
*Riboswitch-cheZ-dT
 
-
*2x cheZ-dT (in case Ribo-cheZ-dT isn't actually there)
 
-
*3x C-EYFP-dT
 
-
*2x GFP-dT
 
-
 
-
Miniprep:
 
-
*Mr. Gene mms6
 
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<li><a href="https://2009.igem.org/Team:Lethbridge">Home</a></li>
 
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===Sat Oct 3===
 
-
Miniprep:
 
-
*sRBS + Lumazine-dT
 
-
*mRBS + N-term FPs
 
-
*C-term FPs + dT
 
-
*pSB + N-term FPs
 
-
*pSB + N-term FPs
 
-
*sRBS + Lumazine-dT
 
-
Restriction:
 
-
 
-
Analytic and Prep:
 
-
*mRBS + N-term FPs
 
-
*C-term FPs + dT
 
-
*pSB + N-term FPs
 
-
*pSB + N-term FPs
 
-
 
-
Analytic only:
 
-
*Mr. Gene mms6
 
-
 
-
Prep only:
 
-
*pBAD-TetR
 
-
 
-
===Oct 5===
 
-
 
-
Gel extraction:
 
-
*sRBS + Lumazine-dT
 
-
*mRBS + N-term FPs
 
-
*C-term FPs + dT
 
-
*pSB + N-term FPs
 
-
*pSB + N-term FPs
 
-
 
-
Ligation:
 
-
*pStrong + Riboswitch-cheZ-dT
 
-
*Riboswitch-GFP-dT
 
-
*pLac + sRBS-Lumazine-dT
 
-
*pBad-TetR + C-term FPs
 
-
*pBad-TetR + N-term FPs
 
-
*pTetR + N-term
 
-
 
-
Transformation:
 
-
*pLac + sRBS-Lumazine-dT
 
-
*pBad-TetR + C-term FPs
 
-
*pBad-TetR + N-term FPs
 
-
*pTetR + N-term ECFP
 
-
*pStrong + Riboswitch-cheZ-dT
 
-
*Riboswitch-GFP-dT
 
-
 
-
Sequencing:
 
-
*sRBS + Lumazine-dT
 
-
*mRBS + N-term FPs
 
-
*C-term FPs + dT
 
-
*pSB + N-term FPs
 
-
*pSB + N-term FPs
 
-
*Mr. Gene mms6
 
-
 
-
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-
 
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===Tues Oct 6===
 
-
Pick Colonies:
 
-
*pLac + sRBS-Lumazine-dT
 
-
*pBad-TetR + C-term FPs
 
-
*pBad-TetR + N-term FPs
 
-
*pTetR + N-term ECFP
 
-
*pStrong + Riboswitch-cheZ-dT
 
-
*Riboswitch-GFP-dT
 
-
 
-
===Wed Oct 7===
 
-
 
-
Miniprep:
 
-
*pLac + sRBS-Lumazine-dT
 
-
*pBad-TetR + C-term FPs
 
-
*pBad-TetR + N-term FPs
 
-
*pTetR + C-term ECFP
 
-
*pTetR + N-term ECFP
 
-
*pStrong + Riboswitch-cheZ-dT
 
-
*Riboswitch-GFP-dT
 
-
 
-
Analytic Restriction:
 
-
*pLac + sRBS-Lumazine-dT
 
-
*pBad-TetR + C-term FPs
 
-
*pBad-TetR + N-term FPs
 
-
*pTetR  + N-term ECFP
 
-
*pStrong + Riboswitch-cheZ-dT
 
-
*Riboswitch-GFP-dT
 
-
 
-
Prep Restriction:
 
-
*Riboswitch-GFP-dT
 
-
<html>
 
-
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-
 
-
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.qmmc .qmdivider{display:block;font-size:1px;border-width:0px;border-style:solid;position:relative;z-index:1;}.qmmc .qmdividery{float:left;width:0px;}.qmmc .qmtitle{display:block;cursor:default;white-space:nowrap;position:relative;z-index:1;}.qmclear {font-size:1px;height:0px;width:0px;clear:left;line-height:0px;display:block;float:none !important;}.qmmc {position:relative;zoom:1;z-index:10;}.qmmc a, .qmmc li {float:left;display:block;white-space:nowrap;position:relative;z-index:1;}.qmmc div a, .qmmc ul a, .qmmc ul li {float:none;}.qmsh div a {float:left;}.qmmc div{visibility:hidden;position:absolute;}.qmmc .qmcbox{cursor:default;display:block;position:relative;z-index:1;}.qmmc .qmcbox a{display:inline;}.qmmc .qmcbox div{float:none;position:static;visibility:inherit;left:auto;}.qmmc li {z-index:auto;}.qmmc ul {left:-10000px;position:absolute;z-index:10;}.qmmc, .qmmc ul {list-style:none;padding:0px;margin:0px;}.qmmc li a {float:none;}.qmmc li:hover>ul{left:auto;}#qm0 ul {top:100%;}#qm0 ul li:hover>ul{top:0px;left:100%;}
 
-
 
-
 
-
/*!!!!!!!!!!! QuickMenu Styles [Please Modify!] !!!!!!!!!!!*/
 
-
 
-
 
-
/* QuickMenu 0 */
 
-
 
-
/*"""""""" (MAIN) Container""""""""*/
 
-
#qm0
 
-
{
 
-
width:auto;
 
-
background-color:transparent;
 
-
}
 
-
 
-
 
-
/*"""""""" (MAIN) Items""""""""*/
 
-
#qm0 a
 
-
{
 
-
padding:5px 60px 3px 0px;
 
-
margin:0px 5px 0px 0px;
 
-
color:#FFD600;
 
-
font-family:Arial;
 
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font-size:12px;
 
-
text-decoration:none;
 
-
font-weight:bold;
 
-
border-width:0px 0px 8px 0px;
 
-
border-style:solid;
 
-
border-color:#FFD600;
 
-
}
 
-
 
-
 
-
/*"""""""" (MAIN) Hover State""""""""*/
 
-
#qm0 a:hover
 
-
{
 
-
color:#3C13AF;
 
-
border-color:#FFAD00;
 
-
}
 
-
 
-
 
-
/*"""""""" (MAIN) Hover State - (duplicated for pure CSS)""""""""*/
 
-
#qm0 li:hover>a
 
-
{
 
-
color:#3C13AF;
 
-
border-color:#FFAD00;
 
-
}
 
-
 
-
 
-
/*"""""""" (MAIN) Parent Items""""""""*/
 
-
#qm0 .qmparent
 
-
{
 
-
background-repeat:no-repeat;
 
-
background-position:95% 50%;
 
-
}
 
-
 
-
 
-
/*"""""""" (MAIN) Active State""""""""*/
 
-
body #qm0 .qmactive, body #qm0 .qmactive:hover
 
-
{
 
-
color:#3C13AF;
 
-
text-decoration:none;
 
-
border-color:#FFAD00;
 
-
}
 
-
 
-
 
-
/*"""""""" (SUB) Container""""""""*/
 
-
#qm0 div, #qm0 ul
 
-
{
 
-
padding:10px 5px 10px 5px;
 
-
background-color:#EEEEEE;
 
-
border-width:1px;
 
-
border-style:solid;
 
-
border-color:#3C13AF;
 
-
}
 
-
 
-
 
-
/*"""""""" (SUB) Items""""""""*/
 
-
#qm0 div a, #qm0 ul a
 
-
{
 
-
padding:3px 5px 3px 5px;
 
-
background-color:transparent;
 
-
color:#230672;
 
-
font-size:11px;
 
-
font-weight:normal;
 
-
border-width:0px;
 
-
border-style:none;
 
-
border-color:#230672;
 
-
}
 
-
 
-
 
-
/*"""""""" (SUB) Hover State""""""""*/
 
-
#qm0 div a:hover
 
-
{
 
-
background-color:#476DD5;
 
-
color:#FFFFFF;
 
-
text-decoration:none;
 
-
}
 
-
 
-
 
-
/*"""""""" (SUB) Hover State - (duplicated for pure CSS)""""""""*/
 
-
#qm0 ul li:hover>a
 
-
{
 
-
background-color:#476DD5;
 
-
color:#FFFFFF;
 
-
text-decoration:none;
 
-
}
 
-
 
-
 
-
/*"""""""" (SUB) Parent Items""""""""*/
 
-
#qm0 div .qmparent, #qm0 ul .qmparent
 
-
{
 
-
background-image:url(qmimages/arrow_0.gif);
 
-
}
 
-
 
-
 
-
/*"""""""" (SUB) Active State""""""""*/
 
-
body #qm0 div .qmactive, body #qm0 div .qmactive:hover
 
-
{
 
-
background-color:#476DD5;
 
-
background-image:url(qmimages/arrow_1.gif);
 
-
color:#FFFFFF;
 
-
}
 
-
 
-
 
-
/*"""""""" Individual Titles""""""""*/
 
-
#qm0 .qmtitle
 
-
{
 
-
margin:2px 5px 5px 5px;
 
-
color:#222222;
 
-
font-family:Arial;
 
-
font-size:11px;
 
-
font-weight:bold;
 
-
}
 
-
 
-
 
-
/*"""""""" Individual Horizontal Dividers""""""""*/
 
-
#qm0 .qmdividerx
 
-
{
 
-
border-top-width:1px;
 
-
margin:4px 5px 4px 5px;
 
-
border-color:#999999;
 
-
}
 
-
 
-
 
-
/*"""""""" Custom Rule""""""""*/
 
-
ul#qm0 ul
 
-
{
 
-
width:100%;
 
-
}
 
-
 
-
 
-
/*"""""""" Custom Rule""""""""*/
 
-
ul#qm0 li:hover > a.qmparent
 
-
{
 
-
background-image:url(qmimages/arrow_1.gif);
 
-
}
 
-
 
-
 
-
</style>
 
-
 
-
<!-- Add-On Core Code (Remove when not using any add-on's) -->
 
-
<style type="text/css">.qmfv{visibility:visible !important;}.qmfh{visibility:hidden !important;}</style><script type="text/javascript">if (!window.qmad){qmad=new Object();qmad.binit="";qmad.bvis="";qmad.bhide="";}</script>
 
-
 
-
<!-- Add-On Settings -->
 
-
<script type="text/JavaScript">
 
-
 
-
/*******  Menu 0 Add-On Settings *******/
 
-
var a = qmad.qm0 = new Object();
 
-
 
-
// Match Widths Add On
 
-
a.mwidths_active = true;
 
-
 
-
// IE Over Select Fix Add On
 
-
a.overselects_active = true;
 
-
 
-
</script>
 
-
 
-
<!-- Core QuickMenu Code -->
 
-
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-
 
-
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-
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-
 
-
<!-- Add-On Code: IE Over Select Fix -->
 
-
<script type="text/javascript">/* <![CDATA[ */if(window.showHelp&&!window.XMLHttpRequest){if(qmad.bvis.indexOf("qm_over_select(b.cdiv);")==-1){qmad.bvis+="qm_over_select(b.cdiv);";qmad.bhide+="qm_over_select(a,1);";}};function qm_over_select(a,hide){var z;if((z=window.qmv)&&(z=z.addons)&&(z=z.over_select)&&!z["on"+qm_index(a)])return;if(!a.settingsid){var v=a;while(!qm_a(v))v=v[qp];a.settingsid=v.id;}var ss=qmad[a.settingsid];if(!ss)return;if(!ss.overselects_active)return;if(!hide&&!a.hasselectfix){var f=document.createElement("IFRAME");f.style.position="absolute";f.style.filter="alpha(opacity=0)";f.src="javascript:false;";f=a.parentNode.appendChild(f);f.frameborder=0;a.hasselectfix=f;}var b=a.hasselectfix;if(b){if(hide)b.style.display="none";else {var oxy=0;if(a.hasshadow&&a.hasshadow.style.visibility=="inherit")oxy=parseInt(ss.shadow_offset);if(!oxy)oxy=0;b.style.width=a.offsetWidth+oxy;b.style.height=a.offsetHeight+oxy;b.style.top=a.style.top;b.style.left=a.style.left;b.style.margin=a.currentStyle.margin;b.style.display="block";}}}/* ]]> */</script>
 
-
</head>
 
-
<body>
 
-
<ul id="qm0" class="qmmc">
 
-
 
-
<li><a href="https://2009.igem.org/Team:Lethbridge">Home</a></li>
 
-
<li><a href="https://2009.igem.org/Team:Lethbridge/Notebook">Top of Page</a></li>
 
-
<li class="qmclear">&nbsp;</li></ul>
 
-
 
-
<!-- Create Menu Settings: (Menu ID, Is Vertical, Show Timer, Hide Timer, On Click (options: 'all' * 'all-always-open' * 'main' * 'lev2'), Right to Left, Horizontal Subs, Flush Left, Flush Top) -->
 
-
<script type="text/javascript">qm_create(0,false,0,500,false,false,false,false,false);</script>
 
-
</body>
 
-
</html>
 
-
 
-
===Thurs Oct 8===
 
-
Gel Extraction:
 
-
*Riboswitch-GFP-dT
 
-
 
-
Ligation:
 
-
*pStrong + Riboswitch-GFP-dT
 
-
 
-
Transformation:
 
-
*pStrong + Riboswitch-GFP-dT
 
-
 
-
Sequencing:
 
-
*pLac-sRBS-Lumazine-dT
 
-
*pBad-TetR-C-term FPs
 
-
*pBad-TetR-N-term FPs
 
-
*pTetR-N-term ECFP
 
-
*pStrong-Riboswitch-cheZ-dT
 
-
*Riboswitch-GFP-dT
 
-
 
-
Maxiprep:
 
-
*pLac-sRBS-Lumazine-dT
 
-
*pBad-TetR-C-term FPs
 
-
*pBad-TetR-N-term FPs
 
-
*pTetR-N-term ECFP
 
-
*pStrong-Riboswitch-GFP-dT
 
-
 
-
Restriction:
 
-
*pLac-sRBS-Lumazine-
 
-
 
-
Analytic and Prep:
 
-
*pBad-TetR-C-term FPs
 
-
*pBad-TetR-N-term FPs
 
-
*pTetR-N-term ECFP
 
-
*C-term CFP (has pTetR)
 
-
 
-
Analytic Only:
 
-
*pStrong-Riboswitch-GFP-dT
 
-
 
-
Fri Oct 9 Overexpression pLac + sRBS-Lumazine-dT
 
-
Gel Extraction pLac-sRBS-Lumazine-dT
 
-
pBad-TetR-C-term FPs
 
-
pBad-TetR-N-term FPs
 
-
pTetR-N-term ECFP
 
-
C-term CFP (has pTetR)
 
-
Motility Media
 
-
===Oct 12===
 
-
Ligation:
 
-
*pLac-sRBS-Lumazine-dT + pBad-TetR-C-term EYFP
 
-
*pLac-sRBS-Lumazine-dT + pBad-TetR-N-term EYFP
 
-
 
-
Transformation:
 
-
*pLac-sRBS-Lumazine-dT + pBad-TetR-C-term EYFP
 
-
*pLac-sRBS-Lumazine-dT + pBad-TetR-N-term EYFP
 
-
 
-
Plate Testing:
 
-
*pStrong-Riboswitch-GFP-dT
 
-
 
-
Motility Test:
 
-
*pStrong + Riboswitch-cheZ-dT
 
-
<html>
 
-
<head>
 
-
 
-
<!--%%%%%%%%%%%% QuickMenu Styles [Keep in head for full validation!] %%%%%%%%%%%-->
 
-
<style type="text/css">
 
-
 
-
 
-
/*!!!!!!!!!!! QuickMenu Core CSS [Do Not Modify!] !!!!!!!!!!!!!*/
 
-
.qmmc .qmdivider{display:block;font-size:1px;border-width:0px;border-style:solid;position:relative;z-index:1;}.qmmc .qmdividery{float:left;width:0px;}.qmmc .qmtitle{display:block;cursor:default;white-space:nowrap;position:relative;z-index:1;}.qmclear {font-size:1px;height:0px;width:0px;clear:left;line-height:0px;display:block;float:none !important;}.qmmc {position:relative;zoom:1;z-index:10;}.qmmc a, .qmmc li {float:left;display:block;white-space:nowrap;position:relative;z-index:1;}.qmmc div a, .qmmc ul a, .qmmc ul li {float:none;}.qmsh div a {float:left;}.qmmc div{visibility:hidden;position:absolute;}.qmmc .qmcbox{cursor:default;display:block;position:relative;z-index:1;}.qmmc .qmcbox a{display:inline;}.qmmc .qmcbox div{float:none;position:static;visibility:inherit;left:auto;}.qmmc li {z-index:auto;}.qmmc ul {left:-10000px;position:absolute;z-index:10;}.qmmc, .qmmc ul {list-style:none;padding:0px;margin:0px;}.qmmc li a {float:none;}.qmmc li:hover>ul{left:auto;}#qm0 ul {top:100%;}#qm0 ul li:hover>ul{top:0px;left:100%;}
 
-
 
-
 
-
/*!!!!!!!!!!! QuickMenu Styles [Please Modify!] !!!!!!!!!!!*/
 
-
 
-
 
-
/* QuickMenu 0 */
 
-
 
-
/*"""""""" (MAIN) Container""""""""*/
 
-
#qm0
 
-
{
 
-
width:auto;
 
-
background-color:transparent;
 
-
}
 
-
 
-
 
-
/*"""""""" (MAIN) Items""""""""*/
 
-
#qm0 a
 
-
{
 
-
padding:5px 60px 3px 0px;
 
-
margin:0px 5px 0px 0px;
 
-
color:#FFD600;
 
-
font-family:Arial;
 
-
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<li><a href="https://2009.igem.org/Team:Lethbridge">Home</a></li>
 
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-
 
-
===Oct 13===
 
-
Pick Colonies
 
-
*pLac-sRBS-Lumazine-dT + pBad-TetR-C-term EYFP
 
-
*pLac-sRBS-Lumazine-dT + pBad-TetR-N-term EYFP
 
-
 
-
Analysis of Plate and Motility Tests
 
-
 
-
===Wed Oct 14===
 
-
Miniprep:
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-C-term EYFP
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-N-term EYFP
 
-
 
-
Restriction:
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-C-term EYFP
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-N-term EYFP
 
-
 
-
===Thurs Oct 15===
 
-
Gel Extraction:
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-C-term EYFP
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-N-term EYFP
 
-
 
-
Ligation:
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-C-term EYFP + C-term CFP
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-N-term EYFP + pTetR-N-term CFP
 
-
 
-
Transformation:
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-C-term EYFP + C-term CFP
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-N-term EYFP + pTetR-N-term CFP
 
-
 
-
Expression Testing:
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-C-term EYFP
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-N-term EYFP
 
-
<html>
 
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<head>
 
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<li><a href="https://2009.igem.org/Team:Lethbridge">Home</a></li>
 
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</html>
 
-
 
-
===Fri Oct 16===
 
-
Pick Colonies:
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-C-term EYFP + C-term CFP
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-N-term EYFP + pTetR-N-term CFP
 
-
 
-
===Sat Oct 17===
 
-
Maxiprep:
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-C-term EYFP + C-term CFP
 
-
*pLac-sRBS-Lumazine-dT-pBad-TetR-N-term EYFP + pTetR-N-term CFP
 
-
<html>
 
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<head>
 
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/*!!!!!!!!!!! QuickMenu Styles [Please Modify!] !!!!!!!!!!!*/
 
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margin:0px 5px 0px 0px;
 
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text-decoration:none;
 
-
font-weight:bold;
 
-
border-width:0px 0px 8px 0px;
 
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border-style:solid;
 
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border-color:#FFD600;
 
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color:#3C13AF;
 
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background-repeat:no-repeat;
 
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color:#3C13AF;
 
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-
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/*"""""""" (SUB) Hover State - (duplicated for pure CSS)""""""""*/
 
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background-color:#476DD5;
 
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#qm0 .qmtitle
 
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<script type="text/JavaScript">
 
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/*******  Menu 0 Add-On Settings *******/
 
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// Match Widths Add On
 
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a.mwidths_active = true;
 
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Revision as of 19:05, 21 October 2009


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