Customers often ask, “Why is the background so high? Why do I have so many wildtype colonies?”
Wildtype colonies result from template DNA that is carried over from the PCR into the final transformation step. To eliminate template DNA, first, be sure that the PCR reaction has worked efficiently. Then, use only 1ul of the PCR reaction in the KLD reaction. This will eliminate plasmid DNA and reduce the number of background colonies.
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