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. 2017 Jun 21;12(6):e0178027. doi: 10.1371/journal.pone.0178027

Fig 2. Addition of a BaseBlocker™ strongly increases SensiScreen® assay specificity.

Fig 2

(A) Normalized real-time PCR amplification plot showing the specificity of SensiScreen® G12S simplex assay in the absence and presence of a wild type blocking BaseBlocker™. (B+C) The inclusion of a BaseBlocker™ (+) impairs amplification of the wild-type allele by 99.95% (B) and increases the difference in threshold cycle (Ct) value between wild-type (WT) and mutant (G12S) DNA samples from -0.7 to >11.2 (C). SensiScreen® KRAS G12S Simplex real-time PCR ±2000 nM of BaseBlocker™ was performed on a Corbett Rotor-Gene 6000 instrument using 50 ng of wild type (WT) human genomic DNA and approximately 500 copies of KRAS G12S template. PCR amplification plot (A) is representative of three independent experiments. Bars (B) represent the mean +S.D. of three independent experiments.