Store: Store at -20°C in the dark.
Code No.:ZB136
Package:50g
Price:EUR(€)1000.00
Description
X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) 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).
Applications:
•Blue/white colony screening assay, distinguishing recombinant colonies (white) among non-recombinant ones (blue), see Protocol for the Blue/White Colony Screening.
•Visualization of expression of the beta-galactosidase reporter gene in transfected eukaryotic cells, see Protocol for the Detection of the Beta-galactosidase Reporter Gene in Transfected Eukaryotic Cells.
•Detection of beta-galactosidase activity in immunological and histochemical procedures.
Quality Control:
Greater than 98% purity confirmed by HPLC.
Functionally tested in blue/white colony screening.
Note
•Preparation of a 20 mg/ml stock solution in dimethylformamide or dimethylsulfoxide is recommended.
•Dimethylformamide dissolves some plastic materials. The direct addition of dimethylformamide containing solution to plastic Petri dishes should be avoided.
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.
ShineGene Bio-Technologies, Inc., located in Shanghai, P.R. China, is a bio-company specialized in providing bio- technical services including Gene Synthesis,Peptide Synthesis,Real Time PCR, Fluorogenic Probe Design,Fluorogenic Probe Label,HRP Enzyme Stabilizer,X-gal,IPTG,ProClin300etc. and supplying reagents such as Hotstart Taq polymerase, dNTPs, qPCR mastermix,one step qPCR kits,First Strand cDNA Synthesis Kits,etc. ShineGene also undertake some OEM or OED orders as well as production task.ShineGene works at the cutting edge of development in PCR, quantitative real-time PCR, transposon and RNA technologies. ShineGenes mission is to provide new tools of excellent quality and extreme purity to the scientific community. Therefore research and development is a continuous process in our company.