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The working principle and application method of IPTG and x-gal in blue-white spot screening

Author:Hunan Hui Bai Yi new materias Co; Ltd Click: Time:2019-12-31 00:00:00

Screening principle of bacterial blue-white spots:

IPTG is a placebo inducer that can induce the production of alpha-galactosidase, and x-gal is the substrate of alpha-galactosidase. Under the action of alpha-galactosidase, x-gal is cut into galactosidase and dark blue substance: 5-bromo-4-indigo, colored substances can make the colony appear blue.


working concentration:

IPTG concentration: 50mg/ml;X-gal concentration: 20mg/ml


Direct coating:

10l of IPTG solution and 20l of x-gal solution were added to the medium, and 50 ~ 100l of sterile water or sterile LB was added at the same time.


prefabricated slab:

When the plate is inverted, the temperature drops to 50℃.250l of IPTG solution and 500 mg per 100mlLB?L of the x-gal solution, after mixing the plate can be made.


attention:

1. X-gal was insoluble in water and was dissolved in dimethylformamide (DMF);

2. X-gal is sensitive to light, and the plate containing x-gal should be kept away from light and used within 1 to 2 weeks;

3. After overnight culture, the blue-white spot is not obvious. The plate can be put into the refrigerator at 4℃.


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E-mail: ivy@hnhbsj.com     

Factory Address:  No.8, Hongjiang Industrial Park, Huaihua, Hunan, China

 

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