Team:UNIPV-Pavia/Methods Materials/Transformation

From 2009.igem.org

(Difference between revisions)
(→Transformation)
(→Transformation)
 
Line 43: Line 43:
*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 class='beuta' align='center' valign='top'  width='300px'>
+
<br></td><td align='center' valign='top'  width='300px'>
<table border='1'>
<table border='1'>

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"