Greater Specificity in Yaku versus Traditional Primer Design in AS-PCR Diagnostic of G85E CFTR mutation in IB3 C

Figure 4: UV illumination of traditional method primers vs. Yaku-method primers PCR product annealing at 49°C. Thermocycling conditions included a five minute initial denaturation at 94^○C with 30 cycles of 30 s at 94^○C, 30 s at 49^○C and 30 s at 72^○C, with a final extension of 7 minutes at 72^○C. Primers were diluted in a 1:10 ratio. Lane 1 contains 1 Kb Plus DNA ladder. Gel electrophoresis was run at 275 volts. Lane 4 contains WT1 with DNA, with Lane 5 serving as the negative control. Lane 7 contains WT2 with DNA; Lane 8 serving as the negative control. Lane 10 contains MT1 with DNA, with Lane 11 serving as the negative control. Lane 13 contains MT2 with DNA; Lane 14 serving as the negative control. Expected bands at 600 base pairs are clearly visible in Lanes 4, 7 and 10, although some smearing is also seen. Lane 13 contains no distinguishable bands.