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Gap ligation approach for sequencing a single uracil lesion in the KRAS sequence. (a) The five-step scheme comprising excision, amplification, and Sanger sequencing of a lesion. (b) PAGE analysis monitoring and quantifying enzyme-catalyzed labeling reactions on the KRAS-U duplex. Reactions were monitored by 32P labeling of the lesion-containing strand, and the gels were quantified by phos-phorimager autoradiography. (c) Percent yield for each enzymatic step.
