KOD -Plus- is based on DNA polymerase from the hyperthermophilic Archaeon Thermococcus kodakaraensis KOD1(1)(2) KOD -Plus- exhibits excellent high PCR fidelity and efficiency. The enzyme solution of KOD -Plus- contains two types of anti-KOD DNA polymerase antibodies that inhibit polymerase and 3'→5' exonuclease activity, thus allowing for Hot Start PCR(3). KOD -Plus- generates blunt-end PCR products, due to 3'→5' exonuclease (proof-reading) activity.
The PCR error ratio of KOD -Plus- is approximately 80 times less than that of Taq DNA polymerase.
Effective for the amplification of GC-rich targets.
Hotstart technology enables highly efficient amplification.
KOD -Plus- enables the following amplifications (maximum): 21 kb from lambda phage DNA, 12 kb from human genomic DNA, and 7 kb from cDNA.
Table 1.Comparison of the mutation frequency of each PCR enzyme.
*PCR fidelity was based on the mutation frequency of PCR products using a positive-selection base assay with the rpsL gene(4).
One unit is defined as the amount of enzyme that will incorporate 10 nmoles of dNTP into an acid insoluble material in 30 min at 75ºC.
*The enzyme solution contains anti-KOD DNA polymerase antibodies that neutralize polymerase and 3'→5' exonuclease activity.
* Do not use dNTPs from other kits or companies.
The GC rich target, lipase gene (Pseudomonas sp.) [GC content = 70%], was amplified using various PCR enzymes. KOD -Plus- could amplify the target gene without additives such as DMSO or PCR enhancer.
Long targets ranging in size from 0.6 to 3.6 kb were amplified using high fidelity PCR enzymes. All targets could be amplified by KOD -Plus-.