10.1371/journal.pone.0183623.g005 Claudia Huber Claudia Huber Andreas Marx Andreas Marx Real time PCR of prothrombin factor II Human gDNA with KOD pol wildtype and mutants. Public Library of Science 2017 DNA polymerases catalyze KOD PCR mismatch extension selectivity DNA polymerases saturation mutagenesis variant 606Q DNA polymerase fidelity mismatch extension selectivity Fidelity Thermococcus kodakaraensis DNA polymerase 606W sequence family B Thermococcus kodakaraensis DNA polymerase 2017-08-23 17:37:06 Figure https://plos.figshare.com/articles/figure/Real_time_PCR_of_prothrombin_factor_II_Human_gDNA_with_KOD_pol_wildtype_and_mutants_/5337376 <p>Real time PCR curves when using a forward primer that terminates opposite the SNP site in either a matched or mismatched fashion, depending on the template. (A) real time PCR curves obtained when using KOD pol wildtype. No discrimination is seen for either of the DNA samples: G-wt (genotype G/G), A-heterozygous (genotype G/A) or A-homozygous (genotype A/A). (B)+(C)+(D) real time PCR curves when using KOD pols R501C, R606Q, and R606W, respectively. Discrimination is seen for the amplification of the A-homozygous (A/A) Factor II template which is severely slowed down when using the KOD mutants. (E) agarose gel electrophoresis analyzing PCR products from (B)-(D). M = marker. PCR products are only visible in samples containing template DNA with at least one allele that results in a matched primer-template duplex (C-primer with G/G and G/A DNA template); the homozygous A/A template does not yield a PCR product.</p>