Team:TU Darmstadt/protocols/Heat Shock Transformation

From 2013.igem.org

(Difference between revisions)
(Created page with "<html> <style type="text/css"> body { margin:0; padding:15px 0 0; background-color: #ececec; background-image:url("https://static.igem.org/mediawiki/2013/1/14/Background_(3000x180_P...")
 
(7 intermediate revisions not shown)
Line 15: Line 15:
{
{
   background-color: white;
   background-color: white;
-
   background-image: url("/wiki/images/7/70/Hintergrund3.jpg");
+
   background-image: url("/wiki/images/5/5c/Hintergrund_Grün(ohne_Header%2C_Logo_und_Schlagschatten).jpg");
   background-attachment: scroll;
   background-attachment: scroll;
   background-position: 50% 0;
   background-position: 50% 0;
Line 26: Line 26:
{
{
     background: white;
     background: white;
-
     background-image: url('/wiki/images/2/2f/Header_(mit_Logo).jpg');
+
     background-image: url("/wiki/images/2/2f/Header_%28mit_Logo%29.jpg");
     margin: 0 auto;
     margin: 0 auto;
     height:250px ;
     height:250px ;
Line 35: Line 35:
     background-repeat: no-repeat;
     background-repeat: no-repeat;
     background-size: 100% 100% ;
     background-size: 100% 100% ;
-
     border-style: none;
+
     border-style: none;;}
-
}
+
#p-logo { display:none;}
#p-logo { display:none;}
Line 42: Line 41:
p {line-height:1.5em; margin:0 0 15px; text-align:justify; color:white;}
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;}
h1 {font-size:1.8em; font-weight:400; margin:0 0 12px;}
 +
 +
 +
#mm_icon1
 +
{
 +
position: absolute;
 +
top: 150px;
 +
left: 30px;
 +
}
 +
 +
#mm_icon2
 +
{
 +
position: absolute;
 +
top: 150px;
 +
left: 350px;
 +
}
 +
 +
#mm_icon3
 +
{
 +
position: absolute;
 +
top: 150px;
 +
left: 670px;
 +
}
 +
 +
#abstracticon1
 +
{
 +
position: absolute;
 +
top: 150px;
 +
left: 30px;
 +
background:white;
 +
filter:alpha(opacity=83); opacity:0.83;
 +
border:1px solid #aaaaaa;
 +
-moz-border-radius:15px;
 +
-khtml-border-radius:15px;
 +
border-radius:15px;
 +
}
 +
 +
#abstracticon2
 +
{
 +
position: absolute;
 +
top: 150px;
 +
left: 350px;
 +
background:white;
 +
filter:alpha(opacity=83); opacity:0.83;
 +
border:1px solid #aaaaaa;
 +
-moz-border-radius:15px;
 +
-khtml-border-radius:15px;
 +
border-radius:15px;
 +
}
 +
 +
 +
 +
#abstracticon3
 +
{
 +
position: absolute;
 +
top: 150px;
 +
left: 670px;
 +
background:white;
 +
filter:alpha(opacity=83); opacity:0.83;
 +
border:1px solid #aaaaaa;
 +
-moz-border-radius:5px;
 +
-khtml-border-radius:15px;
 +
border-radius:15px;
 +
}
 +
 +
#taskbar
 +
{
 +
position:absolute;
 +
top:10px;
 +
left:500px;
 +
z-index: 5;
 +
}
 +
 +
 +
 +
 +
dl.igemTUD2013gelpicture
 +
{
 +
border: 1px solid #000;
 +
background-color: #109f71;
 +
width: 110px;
 +
text-align: center;
 +
padding: 5px 5px 5px 5px;
 +
float: right;
 +
margin: 0 0 0 0;
 +
margin-left:15px
 +
}
 +
 +
.igemTUD2013gelpicture dt
 +
{
 +
font-weight: bold;
 +
background-color: #131210;
 +
color: #959289;
 +
padding: 0 0;
 +
margin-bottom: 10px;
 +
}
 +
 +
.igemTUD2013gelpicture dd img
 +
{
 +
border: 1px solid #000;
 +
width: 100px;
 +
height: 200px;
 +
}
 +
 +
.igemTUD2013gelpicture dd
 +
{
 +
margin: 0;
 +
padding: 5px 5px 5px 5px;
 +
font-size: 100%;
 +
text-align: left;
 +
}
 +
 +
 +
 +
 +
dl.igemTUD2013gelpicture2
 +
{
 +
border: 1px solid #000;
 +
background-color: #109f71;
 +
width: 210px;
 +
text-align: center;
 +
padding: 5px 5px 5px 5px;
 +
float: right;
 +
margin: 0 0 0 0;
 +
margin-left:15px
 +
}
 +
 +
.igemTUD2013gelpicture2 dt
 +
{
 +
font-weight: bold;
 +
background-color: #131210;
 +
color: #959289;
 +
padding: 0 0;
 +
margin-bottom: 10px;
 +
}
 +
 +
.igemTUD2013gelpicture2 dd img
 +
{
 +
border: 1px solid #000;
 +
width: 200px;
 +
height: 200px;
 +
}
 +
 +
.igemTUD2013gelpicture2 dd
 +
{
 +
margin: 0;
 +
padding: 5px 5px 5px 5px;
 +
font-size: 100%;
 +
}
 +
LI.list1 {list-style: none; color:white;}
 +
.blacktext {color:black}
 +
p, ul {
 +
    padding: 0;
 +
    margin: 0;
 +
}
</style>
</style>
-
</html>
 
-
<span style="color:white;">
+
<body>
 +
 
 +
<center>
 +
<!-- central main menu -->
 +
 
 +
<br>
 +
<br>
 +
 
 +
<!-- Taskbar -->
 +
<div id="taskbar">
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt">
 +
<img alt="Home_ausgewählt" src="/wiki/images/e/ef/Darmstadt_green_Home.jpg" width="70" height="30"></a>
 +
 
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/problem">
 +
<img alt="Problem" src="/wiki/images/6/66/Darmstadt_green_Problem.jpg" width="100" height="30"></a>
 +
 
 +
 
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/result">
 +
<img alt="result" src="/wiki/images/2/2c/Darmstadt_green_Result.jpg" width="80" height="30"></a>
 +
 
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/safety">
 +
<img alt="safety" src="/wiki/images/7/7a/Darmstadt_green_Safety.jpg" width="90" height="30"></a>
 +
 
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/team">
 +
<img alt="team" src="/wiki/images/a/a4/Darmstadt_green_Team.jpg" width="70" height="30"></a>
 +
<br>
 +
 
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/strategy">
 +
<img alt="team" src="/wiki/images/a/a0/Darmstadt_green_Strategy.jpg" width="100" height="30"></a>
 +
 
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/humanpractice">
 +
<img alt="team" src="/wiki/images/4/4f/Darmstadt_green_Human_Practice.jpg" width="150" height="30"></a>
 +
 
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/modelling">
 +
<img alt="team" src="/wiki/images/0/06/Darmstadt_green_Modelling.jpg" width="110" height="30"></a>
 +
 
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/labbook">
 +
<img alt="team" src="/wiki/images/4/4c/10._Labbook_(angewählt).jpg" width="90" height="30"></a>
 +
 
 +
</div>
 +
 
 +
 
 +
<br><br><br><br>
 +
<center>
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/labbook">
 +
<font size="8" color="#F0F8FF" face="Arial regular">Lab book |</font>
 +
</a>
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/materials">
 +
<font size="8" color="#F0F8FF" face="Arial regular">Materials |</font>
 +
</a>
 +
<a href="https://2013.igem.org/Team:TU_Darmstadt/protocols">
 +
<font size="8" color="#F0F8FF" face="Arial regular">Protocols</font>
 +
</a>
 +
</center>
 +
 
 +
<!-- Heat Shock Transformation  -->
 +
<center>
 +
<br>
 +
 
 +
<h2><font size="6" color="#F0F8FF" face="Arial regular"> Heat Shock Transformation </font></h2>
 +
<div id="all">
 +
<body>
 +
 
 +
<p text-aligne:left style="margin-left:50px; margin-right:50px"><font size="4.5" color="#F0F8FF" face="Arial regular">
 +
<B> Materials<br></B></font></p>
 +
 
 +
<p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
 +
<br>
 +
<B>Equipment<br></B>
 +
<div align="left" style="margin-left:60px; margin-right:50px">
 +
<ul>
 +
<li class=list1>- Heating bath</li>
 +
<li class=list1>- Incubator</li>
 +
</ul>
 +
</div>
 +
</font>
 +
</p>
 +
 
 +
<p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
 +
<br>
 +
<B>Chemicals & consumables<br></B>
 +
<div align="left" style="margin-left:60px; margin-right:50px">
 +
<ul>
 +
<li class=list1>- Ice</li>
 +
<li class=list1>- Pipets + steril tips</li>
 +
<li class=list1>- <a href="https://2013.igem.org/Team:TU_Darmstadt/materials/Media"> dYT Medium</a></li>
 +
<li class=list1>- LB-Agar-Plates + antibiotics</li>
 +
</ul>
 +
</div>
 +
</font>
 +
</p>
 +
<br>
-
{| style="color:green;background:none;"
+
<p text-aligne:left style="margin-left:50px; margin-right:50px"><font size="4.5" color="#F0F8FF" face="Arial regular">
-
!align="center"|[[Team:TU_Darmstadt/labbook|<span style="color:white;font-size:160%;"> Labbook |</span>]]
+
<B> Procedure<br></B></font></p><br>
-
!align="center"|[[Team:TU_Darmstadt/protocols|<span style="color:white;font-size:160%;"> Protocols |</span> ]]
+
<p text-aligne:left style="margin-left:60px; margin-right:60px"><font size="3" color="#F0F8FF" face="Arial regular">
-
!align="center"|[[Team:TU_Darmstadt/materials|<span style="color:white;font-size:160%;"> Materials |</span>]]
+
<div align="left" style="margin-left:30px; margin-right:50px">
-
|}
+
<ol>
 +
<li>Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.</li>
 +
<li>Add the DNA and incubate the suspension for 5 minutes on ice.</li>
 +
<li>Heat shock is done by incubating the cells for 1 minute at 42°C.</li>
 +
<li>Add 1 mL of DYT medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.</li>
 +
<li>Finally the cells are ready to be crossed out.</li>
 +
</ol>
 +
</div>
 +
</font></p>
 +
<br>

Latest revision as of 00:32, 5 October 2013







Lab book | Materials | Protocols

Heat Shock Transformation

Materials


Equipment

  • - Heating bath
  • - Incubator


Chemicals & consumables

  • - Ice
  • - Pipets + steril tips
  • - dYT Medium
  • - LB-Agar-Plates + antibiotics


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.