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>