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Figure 4 | BMC Biotechnology

Figure 4

From: Enhanced annealing of mismatched oligonucleotides using a novel melting curve assay allows efficient in vitro discrimination and restriction of a single nucleotide polymorphism

Figure 4

S1 nuclease cleavage kinetics on stepwise increasing mismatch 'bubble' templates using optimised annealing conditions. Wild-type (A) and mutant (B) oligonucleotide sets were annealed under specific annealing temperatures determined from the melting curve analysis. Annealed oligonucleotides were incubated with 0.1 U/μl S1 nuclease, and incubated at 20°C for the indicated time. Reactions were stopped by adding 1 μl of 0.5 M EDTA and kept on ice prior to electrophoresis. Data is represented as mean percentage of DNA band intensity compared to uncut template ± SEM (N = 3).

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BMC Biotechnology

ISSN: 1472-6750

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