Team:UNIPV-Pavia/Methods Materials/Transformation

From 2009.igem.org

(Difference between revisions)
(→Transformation)
 
(12 intermediate revisions not shown)
Line 1: Line 1:
-
{{UNIPV-Pavia/StyleCss}}
 
<html>
<html>
-
<head>
+
 
-
<link rel="stylesheet" type="text/css" href="http://bioinfo.unipv.it/igem/templates/SpryAssets/SpryMenuBar.js" title="Stile"/>
+
-
</head>
+
-
<body>
+
<table width="100%">
<table width="100%">
   <tr>
   <tr>
<td align="center">
<td align="center">
-
<img width="100%" height="180px" src="https://static.igem.org/mediawiki/2009/c/c2/UNIPV_title_MandM.jpg" ></img>
+
<img width="100%" height="180px" src="https://static.igem.org/mediawiki/2009/d/de/UNIPV_title_M%26M.jpg" ></img>
</td>
</td>
   </tr>
   </tr>
</table>
</table>
-
</body>
+
 
</html>
</html>
Line 25: Line 21:
     <font face="Pristina" size="50" ><b>Protocols</b></font>
     <font face="Pristina" size="50" ><b>Protocols</b></font>
     </td>
     </td>
 +
    <td width="50px"></td>
</html>
</html>
   </tr>
   </tr>
Line 37: Line 34:
<br>
<br>
<br>
<br>
-
'''Materials needed:'''
+
<table width='80%'><tr><td>
-
*'''LB agar plates with proper antibiotic added incubated at 37°C'''
+
*Put 1-2 µl of DNA resuspension or ligation into thaw TOP10 cells tube (every tube contains approximately 60 µl of competent cells).
-
*'''Thawed Invitrogen TOP10 cells (every tube contains approximately 60 µl of competent cells)'''
+
-
*'''Resuspended DNA'''
+
-
*'''SOC medium'''
+
-
<br>
+
-
*Put 1-2 µl of DNA resuspension or ligation into TOP10 tube.
+
*Incubate on ice for 30 min.
*Incubate on ice for 30 min.
*Heat shock: 42°C for 1 min.
*Heat shock: 42°C for 1 min.
Line 50: Line 42:
*Plate 200 µl of the solution on a proper agar plate.
*Plate 200 µl of the solution on a proper agar plate.
*Incubate overnight at 37°C.
*Incubate overnight at 37°C.
-
<br>
+
<br><br><br><br><br><br>
 +
<br></td><td align='center' valign='top'  width='300px'>
 +
 
 +
<table border='1'>
 +
<tr bgcolor='yellow'><td>INGREDIENTS:
 +
</td></tr>
 +
<tr><td>
 +
<font class='label'>
 +
'''LB agar plates with proper antibiotic'''<hr color='white' width='70%'>
 +
'''Invitrogen TOP10 cells'''<hr color='white' width='70%'>
 +
'''Resuspended DNA'''<hr color='white' width='70%'>
 +
'''SOC medium'''</font>
 +
</td></tr></table><br><br><br><br>
 +
</td></tr></table>

Latest revision as of 10:26, 21 October 2009

EthanolPVanimation.gif



Protocols


Transformation

(estimated time: 3 hours and 30 min + 12-16 hours overnight incubation)

  • Put 1-2 µl of DNA resuspension or ligation into thaw TOP10 cells tube (every tube contains approximately 60 µl of competent cells).
  • Incubate on ice for 30 min.
  • Heat shock: 42°C for 1 min.
  • Put transformed TOP10 tube on ice and then add 250 µl SOC medium.
  • Incubate 1 hour at 37°C, 220 rpm.
  • Plate 200 µl of the solution on a proper agar plate.
  • Incubate overnight at 37°C.








INGREDIENTS:

LB agar plates with proper antibiotic
Invitrogen TOP10 cells
Resuspended DNA

SOC medium





Retrieved from "http://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Transformation"