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IPTG Solution, ready-to-use

 
#R1171 10x1.5 ml (100 mM aqueous solution)

Related Documents (in pdf, ~30-50 KB):

Certificate of Analysis: #R1171
MSDS (English)
MSDS (English-USA)
MSDS (German)

Description
 100 mM IPTG (isopropyl-beta-D-thiogalactopyranoside) is a non-toxic, stable, 0.22 µm membrane-filtered ready-to-use aqueous solution. It is specially designed for direct use in conjunction with X-Gal to determine the lac phenotype in blue/white colony screening.
The IPTG is a highly stable synthetic analog of lactose. It inactivates the lac repressor and induces synthesis of beta-galactosidase, an enzyme that promotes lactose utilization (1).

Applications

Quality Control
Dioxane-free. Functionally tested in blue/white colony screening.

Note
For blue/white screening, 0.1 mM final IPTG concentration in LB (Luria Broth) media is recommended.

Storage
Store at -20°C.

Protocol for the Blue/White Colony Screening

For individual LB (Luria Broth) agar plates:

  1. Pour sterile warm LB agar (about 25 ml) into a Petri dish.
  2. Dry opened LB plates at room temperature under UV light for about 30 min.
  3. Add 40 µl of the X-Gal Solution (20 mg/ml), ready-to-use.
  4. Add 40 µl of 100 mM IPTG Solution, ready-to-use.
  5. Spread evenly on the plate with a sterile spatula.

For batch use, add the following directly per 1 ml of the liquid LB agar (kept at about 50°C):

  1. 1 µl of X-Gal Solution (20 mg/ml), ready-to-use.
  2. 1 µl of 100 mM IPTG Solution, ready-to-use.
  3. Mix well.
  4. Pour 25 ml of prepared LB agar into each Petri dish.
  5. Dry opened LB plates at room temperature under UV light for about 30 min.

Reference

  1. Sambrook, J., Russell, D.W., Molecular Cloning: A Laboratory Manual, the Third edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1.124-1.125, A1.27, 2001.

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Updated vasario 06, 2008 16:15