Beta-glucuronidase protocols

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(Plate Screen)
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==Plate Screen==
==Plate Screen==
===Materials===
===Materials===
-
*Media of choice
+
*Media of choice  
-
*X-gluc stock solution (0.1M = 52.2mg/mL)
+
*Agar
 +
*X-gluc stock solution (50mg/mL)
===Method===
===Method===
-
 
+
#Prepare your liquid media and add the desired amount of agar (usually 1-2%).
 +
#Autoclave the media for the requisite time.
 +
#Add 1.2μL of X-gluc stock solution for every mL of media (e.g. if making 1L of media add 1.2ml of stock solution)
 +
#If desired, add antibiotic. 
 +
#Pour plates.
===Notes===
===Notes===

Revision as of 00:38, 5 July 2011


Contents

Introduction

These are methods to screen for and assay the activity of the common reporter enzyme β-Glucuronidase (GUS) activity. Because the catalytic activity of β-Glucuronidase is very similar to β-Galactosidase (LacZ) these protocols are also very similar to the LacZ protocols.

Plate Screen

Materials

  • Media of choice
  • Agar
  • X-gluc stock solution (50mg/mL)

Method

  1. Prepare your liquid media and add the desired amount of agar (usually 1-2%).
  2. Autoclave the media for the requisite time.
  3. Add 1.2μL of X-gluc stock solution for every mL of media (e.g. if making 1L of media add 1.2ml of stock solution)
  4. If desired, add antibiotic.
  5. Pour plates.

Notes

Assay

Materials

Method

Notes

Staining

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