KLD Enzyme Mix Reaction Protocol (M0554)

1. Prepare a 10 μl reaction as follows:
 PCR Product  1 µl
 KLD Reaction Buffer (2X)  5 µl
 KLD Enzyme MIx (10X)  1 µl
 Nuclease-free Water  3 µl
 Total Volume  10 µl

2. Mix well by pipetting up and down. Incubate at room temperature (25°C) for 5 minutes. Place on ice or store at -20°C.

 3. Transformation: Add 5 µl of the KLD reaction to 50 µl of competent E. coli. If using NEB 5-alpha Competent E. coli (High Efficiency) (NEB #C2987) follow the transformation protocol in the Q5 Site-Directed Mutagenesis Kit (NEB #E0554/ #E0552) manual.