KOD DNA Polymerase
KOD DNA polymerase is cloned from the DNA polymerase gene of Thermococcus kodakarensis, recombinantly expressed in E. coli and purified. Its robust 3′→5′ exonuclease proofreading activity delivers higher fidelity than Pfu DNA Polymerase, approximately 50 times the fidelity of Taq polymerase.
It is also renowned for ultra-fast polymerization speed: 100–138 bp/s, around 5-fold faster than standard Pfu and 2-fold faster than Taq. It yields abundant PCR products in a short time, suitable for high-fidelity amplification of targets up to 6 kb. KOD produces blunt-ended DNA fragments, applicable for gene cloning, heterologous expression and mutation analysis.
Unit Definition
1 U is defined as the amount of enzyme that incorporates 10 nmol dNTPs into acid-insoluble product within 30 min at 75 ℃.
Main Applications
High-fidelity PCR, cloning PCR, blunt-end DNA fragment amplification.
Kit Contents
| Component |
250 U |
500 U |
| KOD DNA Polymerase (5 U/μL) |
50 μL |
100 μL |
| 10× KOD Buffer |
1 mL |
1 mL |
| SuperPure dNTP Mix (10 mM each) |
100 μL |
200 μL |
Storage
Reference 50 μL PCR Reaction System
| Component |
Volume |
Final Concentration |
| Template DNA |
variable |
＜ 0.5 μg |
| Forward Primer (10 μM) |
1 μL |
0.2 μM |
| Reverse Primer (10 μM) |
1 μL |
0.2 μM |
| 10× KOD Buffer |
5 μL |
1 × |
| dNTP Mixture (10 mM each) |
1 μL |
0.2 mM |
| KOD DNA Polymerase |
0.25–0.5 μL |
— |
| ddH₂O |
to 50 μL |
— |
Important Notes
- Add reagents in order: water → other components → KOD enzyme last. Mix thoroughly and spin down briefly before PCR.
- Prepare the reaction mix on ice to avoid degradation of primers and templates by KOD’s exonuclease activity.
- KOD requires high-purity dNTP; the matched SuperPure dNTP Mix is strongly recommended.
- For plasmid / phage templates: recommended template input 5–20 ng. Cycling parameters need adjustment according to fragment size (<1 kb, 1–2 kb, etc.).
The original text is truncated at “Cycling parameters<1kbp target DNA 1-2kbp target”, reserved for you to supplement later.
