Team:UNIPV-Pavia/Methods Materials/Transformation

From 2009.igem.org

(Difference between revisions)
(→Transformation)
Line 37: Line 37:
<br>
<br>
<br>
<br>
-
'''Materials needed:'''
+
<table width='80%'><tr><td>
-
*'''LB agar plates with proper antibiotic added incubated at 37°C'''
+
-
*'''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.
*Put 1-2 µl of DNA resuspension or ligation into TOP10 tube.
*Incubate on ice for 30 min.
*Incubate on ice for 30 min.
Line 50: Line 45:
*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><br><br><br><br>
 +
<br></td><td align='center' valign='bottom' width='300px'>
 +
<div align='center'>
 +
<table border='1'>
 +
<tr><td>
 +
*'''LB agar plates with proper antibiotic added incubated at 37°C'''
 +
*'''Thawed Invitrogen TOP10 cells (every tube contains approximately 60 µl of competent cells)'''
 +
*'''Resuspended DNA'''
 +
*'''SOC medium'''
 +
</td></tr></table></div><br><br><br><br>
 +
</td></tr></table>

Revision as of 11:38, 30 June 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 TOP10 tube.
  • 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.













  • LB agar plates with proper antibiotic added incubated at 37°C
  • Thawed Invitrogen TOP10 cells (every tube contains approximately 60 µl of competent cells)
  • Resuspended DNA
  • SOC medium




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