Team:Washington/HEAT SHOCK CHEM TRANS

From 2013.igem.org

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<html>
 +
 +
<style type="text/css">
 +
.firstHeading {                                                    /* Gets rid of the default iGEM picture*/
 +
    height:2px;
 +
    visibility:hidden;
 +
}
 +
#catlinks {
 +
    display: none;
 +
}
 +
#content {
 +
    border-left-width:0px;
 +
    border-right-width:0px;
 +
    padding: 0px 5px 0px 5px;
 +
    width:965px;                                                    /* Content width*/
 +
}
 +
#menubar {
 +
    background-color:  #FFFFFF;                              /* left menu bar color*/
 +
}
 +
#menubar ul li a {
 +
    color: #545353; }                                            /* menu text color*/
 +
    .right-menu li a {
 +
    color: red; 
 +
    background-color: #FFFFFF;                              /* right menu bar color*/
 +
}
 +
#p-logo { height 1px; overflow:hidden; display: none;
 +
}
 +
#top-section {     
 +
        background-position: center center;
 +
        background-repeat: no-repeat;
 +
        background-color: #FFFFFF;                        /* top header background color*/
 +
        border-width:0px;
 +
        height:10px;                        /* top header height (determines how low content will start to appear)*/
 +
}
 +
#siteSub {
 +
display:none;
 +
}
 +
#search-controls {                                                /* gets rid of the search bar (it was ugly) */
 +
display:none;
 +
margin-top:0px;
 +
}
 +
#contentSub {
 +
display:none;
 +
}
 +
 +
/*Now to set heading and body fonts*/
 +
body {
 +
background:#000000;
 +
background-image: url("https://static.igem.org/mediawiki/2012/1/1b/Gradientback.jpg");
 +
background-position: center;
 +
background-repeat: repeat-y;
 +
}               
 +
h1
 +
{
 +
color:#262626;
 +
text-align:left;
 +
font-family:"Trebuchet";
 +
font-size:25px;
 +
line-height:100%;
 +
border-bottom: none;
 +
padding:0px 0px 0px 0px; 
 +
margin: 0px 0px 0px 0px;
 +
}
 +
h2
 +
{
 +
color:#262626;
 +
text-align:left;
 +
font-family:"Trebuchet";
 +
font-size:20px;
 +
line-height: 100%;
 +
border-bottom: none; 
 +
padding:0px 0px 0px 0px; 
 +
margin: 0px 0px 0px 0x;
 +
}
 +
h3
 +
{
 +
color:#262626;
 +
text-align:left;
 +
font-family:"Trebuchet";
 +
font-size:17px;
 +
line-height: 100%;
 +
border-bottom: none; 
 +
padding:0px 0px 0px 0px; 
 +
margin: 0px 0px 0px 0x;
 +
}
 +
p
 +
{
 +
color:#262626;
 +
text-align: justify;
 +
font-family:"Arial";
 +
font-size:14px;
 +
line-height:100%
 +
border-bottom: none;
 +
padding:0px 0px 0px 0px; 
 +
margin: 0px 0px 0px 0x;
 +
border:0px
 +
}
 +
body {
 +
color:#808080;
 +
line-height: 1;
 +
font-family:"Arial";
 +
font-size:11px; 
 +
}
 +
 +
.whitebox {
 +
  -moz-box-shadow:  0px 0px 0px #ccc;
 +
  -webkit-box-shadow:  0px 0px 0px #ccc;
 +
  box-shadow:  0px 0px 15px #ccc;
 +
  background-color: #FFFFFF;
 +
  -moz-border-radius: 1px;
 +
  border-radius: 15px;
 +
  margin-top: 20px;
 +
  padding-top: 20px;
 +
  padding-left: 20px;
 +
  padding-right: 20px;
 +
  padding-bottom: 20px;
 +
  margin-left: 15px;
 +
  margin-right: 25px;
 +
}
 +
.thumbnail
 +
{
 +
margin-bottom:0px;
 +
padding-bottom:0px;
 +
position:relative;
 +
}
 +
 +
#igemuwmenu, #igemuwmenu ul {
 +
margin: 0;
 +
padding: 0;
 +
list-style: none;
 +
}
 +
 +
#igemuwmenu {
 +
width: 820px;
 +
margin: 10px auto;
 +
border: 1px solid #222;
 +
background-color: #111;
 +
background-image: -moz-linear-gradient(#444, #111);
 +
background-image: -webkit-gradient(linear, left top, left bottom, from(#444), to(#111));
 +
background-image: -webkit-linear-gradient(#444, #111);
 +
background-image: -o-linear-gradient(#444, #111);
 +
background-image: -ms-linear-gradient(#444, #111);
 +
background-image: linear-gradient(#444, #111);
 +
-moz-border-radius: 6px;
 +
-webkit-border-radius: 6px;
 +
border-radius: 6px;
 +
-moz-box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
 +
-webkit-box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
 +
box-shadow: 0 1px 1px #777, 0 1px 0 #666 inset;
 +
                display: inline-block;
 +
                white-space: nowrap;
 +
}
 +
 +
#igemuwmenu:before,
 +
#igemuwmenu:after {
 +
content: "";
 +
display: table;
 +
}
 +
 +
#igemuwmenu:after {
 +
clear: both;
 +
}
 +
 +
#igemuwmenu {
 +
zoom:1;
 +
}
 +
 +
#igemuwmenu li {
 +
border-right: 1px solid #222;
 +
-moz-box-shadow: 1px 0 0 #444;
 +
-webkit-box-shadow: 1px 0 0 #444;
 +
box-shadow: 1px 0 0 #444;
 +
position: relative;
 +
                display: inline-block;
 +
                white-space: nowrap;
 +
}
 +
 +
#igemuwmenu a {
 +
padding: 12px 20px;
 +
color: #999;
 +
text-transform: uppercase;
 +
font: bold 11.4px Arial, Helvetica;
 +
text-decoration: none;
 +
text-shadow: 0 1px 0 #000;
 +
                display: inline-block;
 +
                white-block: nowrap;
 +
}
 +
       
 +
    #igemuwmenu li:hover > a {
 +
color: #fafafa;
 +
}
 +
 +
*html #igemuwmenu li a:hover { /* IE6 only */
 +
color: #fafafa;
 +
}
 +
 +
#igemuwmenu ul {
 +
margin: 20px 0 0 0;
 +
_margin: 0; /*IE6 only*/
 +
opacity: 0;
 +
visibility: hidden;
 +
position: absolute;
 +
top: 38px;
 +
left: 0;
 +
z-index: 1;   
 +
background: #444;
 +
background: -moz-linear-gradient(#444, #111);
 +
background-image: -webkit-gradient(linear, left top, left bottom, from(#444), to(#111));
 +
background: -webkit-linear-gradient(#444, #111);   
 +
background: -o-linear-gradient(#444, #111);
 +
background: -ms-linear-gradient(#444, #111);
 +
background: linear-gradient(#444, #111);
 +
-moz-box-shadow: 0 -1px rgba(255,255,255,.3);
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-webkit-box-shadow: 0 -1px 0 rgba(255,255,255,.3);
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box-shadow: 0 -1px 0 rgba(255,255,255,.3);
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-moz-border-radius: 3px;
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-webkit-border-radius: 3px;
 +
border-radius: 3px;
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-webkit-transition: all .2s ease-in-out;
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-moz-transition: all .2s ease-in-out;
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-ms-transition: all .2s ease-in-out;
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-o-transition: all .2s ease-in-out;
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transition: all .2s ease-in-out; 
 +
}
 +
 +
#igemuwmenu li:hover > ul {
 +
opacity: 1;
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visibility: visible;
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margin: 0;
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}
 +
 +
#igemuwmenu ul ul {
 +
top: 0;
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left: 150px;
 +
margin: 0 0 0 20px;
 +
_margin: 0; /*IE6 only*/
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-moz-box-shadow: -1px 0 0 rgba(255,255,255,.3);
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-webkit-box-shadow: -1px 0 0 rgba(255,255,255,.3);
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box-shadow: -1px 0 0 rgba(255,255,255,.3);
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}
 +
 +
#igemuwmenu ul li {
 +
float: none;
 +
display: block;
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border: 0;
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_line-height: 0; /*IE6 only*/
 +
-moz-box-shadow: 0 1px 0 #111, 0 2px 0 #666;
 +
-webkit-box-shadow: 0 1px 0 #111, 0 2px 0 #666;
 +
box-shadow: 0 1px 0 #111, 0 2px 0 #666;
 +
}
 +
#igemuwmenu ul li.active {
 +
float: none;
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display: block;
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border: 0;
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_line-height: 0; /*IE6 only*/
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-moz-box-shadow: 0 1px 0 #111, 0 2px 0 #666;
 +
-webkit-box-shadow: 0 1px 0 #111, 0 2px 0 #666;
 +
box-shadow: 0 1px 0 #111, 0 2px 0 #666;
 +
}
 +
 +
 +
#igemuwmenu ul li:last-child { 
 +
-moz-box-shadow: none;
 +
-webkit-box-shadow: none;
 +
box-shadow: none;   
 +
}
 +
 +
#igemuwmenu ul a {   
 +
padding: 10px;
 +
width: 130px;
 +
_height: 10px; /*IE6 only*/
 +
display: block;
 +
white-space: nowrap;
 +
float: none;
 +
text-transform: none;
 +
}
 +
     
 +
        #igemuwmenu ul a.selected {   
 +
padding: 10px;
 +
width: 130px;
 +
_height: 10px; /*IE6 only*/
 +
display: block;
 +
white-space: nowrap;
 +
float: none;
 +
text-transform: none;
 +
}
 +
 +
#igemuwmenu ul a:hover {
 +
background-color: #785D84;
 +
background-image: -moz-linear-gradient(#B6A0BA,  #785D84);
 +
background-image: -webkit-gradient(linear, left top, left bottom, from(#B6A0BA), to(#785D84));
 +
background-image: -webkit-linear-gradient(#B6A0BA, #785D84);
 +
background-image: -o-linear-gradient(#B6A0BA, #785D84);
 +
background-image: -ms-linear-gradient(#B6A0BA, #785D84);
 +
background-image: linear-gradient(#B6A0BA, #785D84);
 +
}
 +
 +
#igemuwmenu ul li:first-child > a {
 +
-moz-border-radius: 3px 3px 0 0;
 +
-webkit-border-radius: 3px 3px 0 0;
 +
border-radius: 3px 3px 0 0;
 +
}
 +
 +
#igemuwmenu ul li:first-child > a:after {
 +
content: '';
 +
position: absolute;
 +
left: 40px;
 +
top: -6px;
 +
border-left: 6px solid transparent;
 +
border-right: 6px solid transparent;
 +
border-bottom: 6px solid #444;
 +
}
 +
 +
#igemuwmenu ul ul li:first-child a:after {
 +
left: -6px;
 +
top: 50%;
 +
margin-top: -6px;
 +
border-left: 0;
 +
border-bottom: 6px solid transparent;
 +
border-top: 6px solid transparent;
 +
border-right: 6px solid #3b3b3b;
 +
}
 +
 +
#igemuwmenu ul li:first-child a:hover:after {
 +
border-bottom-color: #B6A0BA;
 +
}
 +
 +
#igemuwmenu ul ul li:first-child a:hover:after {
 +
border-right-color: #0299d3;
 +
border-bottom-color: transparent;
 +
}
 +
 +
#igemuwmenu ul li:last-child > a {
 +
-moz-border-radius: 0 0 3px 3px;
 +
-webkit-border-radius: 0 0 3px 3px;
 +
border-radius: 0 0 3px 3px;
 +
}
 +
 +
/* Mobile */
 +
#igemuwmenu-trigger {
 +
display: none;
 +
}
 +
 +
@media screen and (max-width: 600px) {
 +
 +
/* nav-wrap */
 +
#igemuwmenu-wrap {
 +
position: relative;
 +
}
 +
 +
#igemuwmenu-wrap * {
 +
-moz-box-sizing: border-box;
 +
-webkit-box-sizing: border-box;
 +
box-sizing: border-box;
 +
}
 +
 +
/* menu icon */
 +
#igemuwmenu-trigger {
 +
display: block; /* show menu icon */
 +
height: 40px;
 +
line-height: 40px;
 +
cursor: pointer;
 +
padding: 0 0 0 35px;
 +
border: 1px solid #222;
 +
color: #fafafa;
 +
font-weight: bold;
 +
background-color: #111;
 +
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==''''''==
==''''''==

Revision as of 06:33, 7 September 2013


'

Heat shock protocol (approximate time 1hrs)

  1. Remove and thaw (on ice) chemically competent cells, either 1.5ml tube or PCR tube.
    1. 1.5mL tube aliquots of cells contain approx. 205uL cells
    2. Pcr tubes aliquots contain 50uls
  2. Add 1-5µl DNA to 40-50µl of competent cells.
  3. ncubate on ice for 5 minutes (for higher transformation efficiency, do 30 min)
  4. Heat shock cells for 45 seconds at 42°C
  5. Let cells stand on ice for 2 minutes (or longer)
  6. Add 200uL2-1mL of LB and shake for 30 min at 37°C (for higher transformation efficiency, do 1 hr). This is called recovery.
  7. Plate 200uL cells on appropriate antibiotic plate1 and grow O/N in 37°C incubator

  1. Preheat plates in 37C before for better results (labeling and cell absorption).
  2. If using 300ul pcr tubes, omit shaking, incubate 37C.

NOTE: Do not use XL10 Gold Cells with Chlor Plates