IGEM:Harvard/2006/TOPO Cloning


 * After purifying the PCR product, add Taq polymerase buffer and dATP to their working concentrations, and 0.5 unit of Taq polymerase. Incubate the reaction for 10-15 minutes at 72°C.
 * To prepare the cloning reaction combine the following:
 * 0.5 to 4 uL PCR product
 * 1 uL salt solution
 * dilute to a total of 5 uL with water
 * 1 uL TOPO vector
 * Mix gently and incubate at room temperature for 5 minutes.
 * Put the reaction on ice and proceed to the standard TOP10 transformation protocol.