Team:UNIPV-Pavia/Methods Materials/XGal

From 2009.igem.org

(Difference between revisions)
Line 40: Line 40:
*'''40 mg/ml X-gal in DMF'''
*'''40 mg/ml X-gal in DMF'''
*'''SOC medium'''
*'''SOC medium'''
-
*'''pre-warmed LB agar plate'''
+
*'''Pre-warmed LB agar plate'''
-
*'''face mask'''
+
*'''Face mask'''
<br>
<br>

Revision as of 23:04, 23 June 2009

EthanolPVanimation.gif



Protocols


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

(Estimated time: 20 min)

Materials needed:

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


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