Fig. 1
From: Development of a Facile Fluorescent Assay for the Detection of 80 Mutations Within the p53 Gene
Preparation of synthetic mutated templates. The method is based on the method of Higuchi et al. (37). In the first round, two fragments are created in separate reactions, each harboring the primer-induced mutation. In the second round, overlapping strands from the two fragments hybridize and produce a cassette with the mutation in the center. Nested primers, also in the second round, then amplify the desired product.