X-Gal Solution, ready-to-use
#R0941 10ml Related Documents (in pdf, ~30-50 KB):
Certificate of Analysis: #R0941
MSDS (English)
MSDS (English-USA)
MSDS (German)Description
X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galacto-pyranoside) is a non-toxic, ready-to-use, stable, 0.22 µm membrane filtered solution of a special formulation. It is designed for direct use in conjunction with IPTG. The X-Gal is an inert chromogenic substrate for beta-galactosidase, an enzyme that promotes lactose utilization. Beta-galactosidase hydrolyzes X-Gal into a colorless galactose and 4-chloro-3-brom-indigo, which forms an intense blue precipitate. Induction of the lacZ gene with IPTG leads to the hydrolysis of X-Gal and to the development of blue colonies (see the scheme below).
Application
Blue/white colony screening assay, distinguishing recombinant colonies (white) among non-recombinant ones (blue) (1), see protocol below.Quality Control
Functionally tested in blue/white colony screening.Concentration
20 mg/mlStorage
Stable for at least 12 months at +4°C or at room temperature in the original dark vial.Protocol for the Blue/White Colony ScreeningFor individual LB (Luria Broth) agar plates:
- Pour sterile warm LB agar (about 25 ml) into a Petri dish.
- Dry opened LB plates at room temperature under UV light for about 30 min.
- Add 40 µl of the X-Gal Solution (20 mg/ml), ready-to-use.
- Add 40 µl of 100 mM IPTG Solution, ready-to-use.
- 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 µl of X-Gal Solution (20 mg/ml), ready-to-use.
- 1 µl of 100 mM IPTG Solution, ready-to-use.
- Mix well.
- Pour 25 ml of prepared LB agar into each Petri dish.
- Dry opened LB plates at room temperature under UV light for about 30 min.
Sambrook, J., Russell, D.W., Molecular Cloning: A Laboratory Manual, the third edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1.10, 1.123-1.125, 1.127, 1.150, 2.30, 16.13, 17.97, 17.99-1.125,A1.27, 2001.
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Updated vasario 27, 2008 12:36