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|- Use 20–25 ng of the target plasmid per 50 µL of mutagenesis reaction as a starting point.|
|- Create a fresh dilution of 25X SAM from the kit-supplied 200X SAM in sterile, distilled water each time you perform the mutagenesis procedure
- 25X SAM is not stable, and loses activity within a few hours after
|- Only use 1 μl of PCR product in the KLD reaction. Carrying too much PCR product forward can decrease transformation efficiency.
- Only use 5 μl of the KLD reaction in the transformation. If more KLD reaction is added, a buffer exchange step, such as PCR
purification, should be included prior to transformation.
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