In the Blue/White Cloning of a DNA Fragment Laboratory Kit for biotechnology, complete a litigation, a transformation, and an assay, which is based upon the blue/white colony screening protocol.
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Blue/white screening is one of the ways scientists determine if cloning has been successful. In this kit, students complete a ligation, a transformation, and an assay, which is based upon the blue/white colony screening protocol. If the gene that expresses β-galactosidase is interrupted by the cloning plasmid, then white colonies will grow because the enzyme is not able to cleave the X-Gal indicator. If the β-galactosidase gene is not cut by the plasmid, then the X-Gal is cleaved into galactose and a blue indigo compound. So, if the colonies are blue, then the insertion failed. Comprehensive Background section explains each step in the process. Three convenient halt points are indicated in the procedure so the experiment can be performed over several days in three 60–70 minute sections. Complete for 5 groups of students. Kit includes the DNA fragment, linear plasmid, T4 DNA Ligase, competent cells X-Gal, and all the necessary reagents. An incubator, two waterbaths, a shaking incubator or shaking waterbath, microwave or hot plate, automatic micropipet and tips, spectrophotometer, balance, centrifuge, and microcentrifuge are all required and available separately. Live Material: Redemption coupon included to obtain live material. Please allow 2–3 weeks to receive live material after sending in the redemption coupon.