Team:UNIPV-Pavia/Methods Materials/Transformation

(Difference between revisions)

Latest revision as of 10:26, 21 October 2009

Protocols

(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

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 *Incubate overnight at 37°C.
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-<br></td><td class='transformation' align='center' valign='bottom' width='300px'>
+<br></td><td align='center' valign='top'  width='300px'>
-<table class='in_transformation' border='1'>
+<table border='1'>
+<tr bgcolor='yellow'><td>INGREDIENTS:
+</td></tr>
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-*'''LB agar plates with proper antibiotic'''
+<font class='label'>
-*'''Invitrogen TOP10 cells'''
+'''LB agar plates with proper antibiotic'''<hr color='white' width='70%'>
-*'''Resuspended DNA'''
+'''Invitrogen TOP10 cells'''<hr color='white' width='70%'>
-*'''SOC medium'''
+'''Resuspended DNA'''<hr color='white' width='70%'>
-</td></tr></table></div><br><br><br><br>
+'''SOC medium'''</font>
+</td></tr></table><br><br><br><br>
 </td></tr></table>