Team:TU Darmstadt/protocols/Heat Shock Transformation

From 2013.igem.org

(Difference between revisions)
Line 2: Line 2:
<style type="text/css">
<style type="text/css">
-
body  
+
body
{
{
margin:0;  
margin:0;  
Line 43: Line 43:
h1 {font-size:1.8em; font-weight:400; margin:0 0 12px;}
h1 {font-size:1.8em; font-weight:400; margin:0 0 12px;}
-
</style>
+
#mm_icon1
-
</html>
+
{
 +
position: absolute;
 +
top: 240px;
 +
left: 200px;
 +
}
-
<span style="color:white;">
+
#mm_icon2
 +
{
 +
position: relative;
 +
top: 170px;
 +
left: 350px;
 +
}
-
{| style="color:green;background:none;"
+
#mm_icon3
-
!align="center"|[[Team:TU_Darmstadt/labbook|<span style="color:white;font-size:160%;"> Labbook |</span>]]
+
{
-
!align="center"|[[Team:TU_Darmstadt/protocols|<span style="color:white;font-size:160%;"> Protocols |</span> ]]
+
position: absolute;
-
!align="center"|[[Team:TU_Darmstadt/materials|<span style="color:white;font-size:160%;"> Materials |</span>]]
+
top: 150px;  
-
|}
+
left: 670px;
-
<html>
+
}
-
 
+
#abstracticon1
-
<style type="text/css">
+
-
body
+
{
{
-
margin:0;  
+
position: absolute;
-
padding:15px 0 0;
+
top: 260px;  
-
background-color: #ececec;
+
left: 230px;
-
background-image:url("https://static.igem.org/mediawiki/2013/1/14/Background_(3000x180_Pixel).jpg");
+
background:white;
-
background-repeat:repeat;
+
filter:alpha(opacity=83); opacity:0.83;
 +
border:1px solid #aaaaaa;
 +
-moz-border-radius:15px;
 +
-khtml-border-radius:15px;
 +
border-radius:15px;
}
}
-
/* Inner Environment (origin item) */
+
#abstracticon2
-
#content
+
{
{
-
  background-color: white;
+
position: absolute;
-
  background-image: url("/wiki/images/7/70/Hintergrund3.jpg");
+
top: 670px;
-
  background-attachment: scroll;
+
left: 350px;
-
  background-position: 50% 0;
+
background:white;
-
  background-repeat: none;
+
filter:alpha(opacity=83); opacity:0.83;
-
  overflow: hidden;
+
border:1px solid #aaaaaa;
-
  border-style: none;
+
-moz-border-radius:15px;
 +
-khtml-border-radius:15px;
 +
border-radius:15px;
}
}
-
 
+
#abstracticon3
-
#top-section
+
{
{
-
    background: white;
+
position: absolute;
-
    background-image: url('/wiki/images/2/2f/Header_(mit_Logo).jpg');
+
top: 150px;  
-
    margin: 0 auto;
+
left: 700px;
-
    height:250px ;
+
background:white;
-
    text-align: center;
+
filter:alpha(opacity=83); opacity:0.83;
-
    z-index: 1;
+
border:1px solid #aaaaaa;
-
    position: relative;
+
-moz-border-radius:5px;
-
    top: 0;
+
-khtml-border-radius:15px;
-
    background-repeat: no-repeat;
+
border-radius:15px;
-
    background-size: 100% 100% ;
+
}
-
    border-style: none;;}
+
-
#p-logo { display:none;}
 
-
 
-
p {line-height:1.5em; margin:0 0 15px; text-align:justify; color:white;}
 
-
h1 {font-size:1.8em; font-weight:400; margin:0 0 12px;}
 
#taskbar
#taskbar
Line 105: Line 111:
z-index: 5;
z-index: 5;
}
}
-
 
</style>
</style>
 +
<!-- Taskbar -->
<!-- Taskbar -->
<div id="taskbar">
<div id="taskbar">
Line 140: Line 146:
<a href="https://2013.igem.org/Team:TU_Darmstadt/labbook">
<a href="https://2013.igem.org/Team:TU_Darmstadt/labbook">
<img alt="team" src="/wiki/images/f/f3/Darmstadt_green_Labbook.jpg" width="90" height="30"></a>
<img alt="team" src="/wiki/images/f/f3/Darmstadt_green_Labbook.jpg" width="90" height="30"></a>
-
 
-
 
</div>
</div>

Revision as of 15:28, 4 October 2013





Heat Shock Transformation

Materials
Ice
Heatingbad
Pipets + steril tips
DYT-Media
LB-Agar-Plates + antibiotics
Incubator

Procedure
1. Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice
2. Add the DNA and incubate the suspension for 5 minutes on ice
3. Heat shock is done by incubating the cells for 1 minute at 42°C
4. Add 1 mL of DYT medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance
5. Finally the cells are ready to be crossed out