Team:UNIPV-Pavia/Methods Materials

From 2009.igem.org

Revision as of 14:39, 22 June 2009 by Susanna (Talk | contribs)

EthanolPVanimation.gif

Protocols

<a href="https://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Lb"> LB medium preparation </a>


Gycerol stock

  • Mix 750 ul from the 5 ml sample of overnight incubated bacteria with 250 ul of 80% glycerol
  • Leave at -20°C for one day
  • Move to -80°C the day after

X-Gal Staining Protocol for beta Galactosidase (blue/white screening) 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
  • Remember to use dark conditions
  • 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"