Team:UNIPV-Pavia/Methods Materials/XGal

From 2009.igem.org

(Difference between revisions)
m
(→X-Gal staining protocol for beta galactosidase (blue/white screening))
 
Line 41: Line 41:
*Result: blue colonies express LacZ, while white colonies don't.
*Result: blue colonies express LacZ, while white colonies don't.
<br><br><br><br><br><br><br><br>
<br><br><br><br><br><br><br><br>
-
<br></td><td class='xgal' align='center' valign='bottom' width='300px'>
+
<br></td><td align='center' valign='top' width='300px'>
-
<table class='in_xgal' border='1'>
+
 
 +
<table border='1'>
 +
<tr bgcolor='yellow'><td>INGREDIENTS:
 +
</td></tr>
<tr><td>
<tr><td>
<font class='label'>
<font class='label'>

Latest revision as of 10:27, 21 October 2009

EthanolPVanimation.gif



Protocols


X-Gal staining protocol for beta galactosidase (blue/white screening)

(Estimated time: 20 min)

  • The principle is that X-Gal (5-bromo-4-chloro-3-indolyl-b-D-galactopyranoside) turns blue when reacts with beta-galactosidase.
  • Mix 20 ul X-Gal 20 mg/ml and 80 ul SOC and spread on LB+amp plates (X-Gal and DMF are toxic, use face-mask for your safety!!! X-Gal is light-sensitive, remember to keep it in the dark, when possible)
  • Incubate with 200 ul bacteria
  • Result: blue colonies express LacZ, while white colonies don't.










INGREDIENTS:

40 mg/ml X-gal in DMF
SOC medium
Pre-warmed LB agar plate

Face mask





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