IGEM:Harvard/2006/TOPO Cloning
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- 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.
