The ligation and transformation module is an integral component of the Cloning and Sequencing Explorer Series. Use this module to directly ligate PCR products into the pJET1.2 plasmid vector and immediately transform freshly prepared competent bacteria with the ligation reaction. Competent cells are then plated directly on warm agar plates and incubated overnight at 37°C.
Once transformed bacteria have been propagated, plasmid DNA can be isolated and digested with BglII enzyme. The BglII restriction sites located on either side of the pJET1.2 cloning site allow students to determine if their PCR product was successfully ligated.
This module may also be purchased separately and used to clone any PCR product. The entire ligation and transformation protocol takes approximately 2 hours to complete and does not require commercial competent cells, a refrigerated microcentrifuge, or a –80°C freezer, making it the method of choice for educators worldwide.
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