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Comparing the speed and product yield of 7 high fidelity DNA Polymerases
The  three Rs of PCR: Rapid, Robust, Reliable

KOD is Rapid
KOD yields more product in fewer cycles compared to other PCR enzymes

The data below show the speed and yield of KOD and KOD Hot Start DNA Polymerases compared to five other high fidelity thermophilic DNA polymerases. In addition to a low mutation frequency, the fast extension rate and high processivity of KOD result in higher yields of full-length product in fewer reaction cycles. Combined, these make KOD Hot Start DNA Polymerase the PCR enzyme of choice for structural proteomics studies.

All 7 enzymes were tested in 4 different cycling protocols, which encompass the manufacutrer's recommended cycling conditions

 
Cycling profile A
PCR samples (5 µl) were taken after 23, 25, 27, and 29 cycles and assayed on 1.4% agarose/TAE gels. Lanes indicated the enzyme used for the reaction. Cycling profiles are defined in the table above.

 
For cycles B, C, and D see the full newsletter article

KOD is Robust
KOD for 2-step PCR

• KOD enzymes give high yields in a time-saving 2-step protocol comparable to the 3-step protocols (Compare to Figure 4 above).

PCR results from 7 high fidelity enzymes using a 2-step cycling profile
PCR samples were removed after 23, 25, 27, and 29 cycles of a 2-step protocol and 5 µl were assayed on 1.4% agarose/TAE gels. Lanes indicated the enzymes used for the reaction.

KOD is Reliable
KOD application: screening cDNA libraries

KOD Hot Start DNA Polymerase was used in a 2-step cycling profile to screen clones from the T7Select® Human cDNA Library (Cat. No. 70637). Of the 50 clones screened, KOD successfully amplified 49 inserts. Amplicons ranged in size from ~250–1800 bp.

KOD Hot Start amplification of T7Select® human cDNA Library
Reactions were cycled with an initial denaturation at 95°C for 2 min and 25 cycles of 95°C for 20 s, 68°C for 25 s.

• Elongation rate is 5 times higher than Pfu DNA Polymerase (3).

• Processivity is 10 to 15 times higher than Pfu DNA polymerase (3).

• Highest yields of amplified human genomic DNA was obtained with KOD Hot Start DNA Polymerase (refer to Figure 3 below). Both PfuTurbo and Pfu Ultra high-fidelity DNA polymerases generated lower yields of products under the same conditions.

• Lower PCR mutation frequency. When the fidelity of replication was measured as the mutation frequency in PCR products using a modified rpsL⁺ fidelity assay it was found that KOD Hot Start and Pfu Ultra demonstrated comparable mutation frequencies, while PfuTurbo exhibited a two-fold higher mutation frequency. (refer to Figure 2).