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.