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. Author manuscript; available in PMC: 2019 Dec 1.
Published in final edited form as: Nat Protoc. 2019 May 10;14(6):1863–1883. doi: 10.1038/s41596-019-0165-3

Figure 4. Quikchange strategy to clone iFKBP, uniRapR, or LOV2 into a target gene.

Figure 4

In Step 1, forward and reverse primers are used in PCR to generate megaprimers consisting of a fragment aligning to the target gene and a flanking region aligned to the inserted sequence. In Step 2, Quikchange PCR is performed using the megaprimers and target plasmid. F and R denote forward and reverse primers. The blue region indicates the target gene, and the blue dots indicate synthesized DNA with Quikchange PCR. The insertion sites of kinases, GEFs and GTPases are at the bottom.