300- Blue/White Cloning of a DNA Fragment and Assay of β-galactosidase
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Price: $518 + GST
Model Number: 300
For 5 Lab Groups.
LigatioTransformation of E.coli with Plasmid pBR322n 70 minutes.
Transformation 60 minutes.
Plating 5 minutes.
Incubation overnight
Transformation 15 minutes.
When DNA is subcloned in the pUC polylinker region, ß-galactosidase production is interrupted, resulting in the inability of cells to hydrolyse X-Gal. This results in the production of white colonies amongst a background of blue colonies. This experiment provides a DNA fragment together with a linear plasmid and T4 DNA Ligase. Following the ligation to synthesize the recombinant plasmid, competent E. coli cells are transformed and the number of recombinant antibiotic resistant white
and blue colonies are counted. ß-galactosidase activity is assayed from blue and white bacterial cells. This experiment can be broken down into three modules: ligation, transformation, and assay of ß-galactosidase.
Kit includes:
- Instructions
- Linearized pUC plasmid & DNA fragment
- T4 Ligase
- Bacterial LyphoCells™ for transformation
- Reconstitution buffer
- XGal in solvent
- IPTG
- Calcium chloride
- Aantibiotic
- ReadyPour™ Luria Broth Agar
- Luria broth media for recovery
- Growth media
- Assay components
- Plastic supplies
Requirements:
- Incubation oven
- Waterbath
- Shaking incubator or shaking waterbath (or be prepared to swirl cultures from time to time)
- Automatic micropipet and tips
- Spectrophotometer
- Centrifuge
- Microcentrifuge
- Ice
