P404 Detailed protocol for cloning included. Shipped on dry ice via overnight delivery. PCR-TRAP cloning system. A schematic diagram of the PCR-TRAP cloning vector before and after cloning of a PCR product. PCR-TRAP Cloning Vector ligated with a 450 bp PCR amplified LacZ gene was transformed into E. coli and plated on a tetracycline plate.
Blue colonies are positive for insert, while white colonies indicate background colonies.
Colony PCR analysis of the 450 bp LacZ inserts cloned into the PCR-TRAP vector. Lane one is a 450 bp LacZ PCR fragment before cloning. Lane 2 is a 100 bp DNA ladder as a MW standard. TetR colonies that were checked for insert by colony PCR using a pair of primers located 60 bp away from either side of the cloning site. All positive clones will give a PCR product 120 bp bigger than the actual insert before cloning.