KOD DNA Polymerase |
| Katalog-Nr.GE10004 |
DNA polymerase from the extremely heat-resistant archaeon Thermococcus kodakaraensis (KOD) is a new high-fidelity, fast-heat-resistant DNA polymerase.
Products are for research use only. Not for human use. We do not sell to patients.
Sample solution is provided at 25 µL, 10mM.
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DNA polymerase from the extremely heat-resistant archaeon Thermococcus kodakaraensis (KOD) is a new high-fidelity, fast-heat-resistant DNA polymerase. The amplification speed of this enzyme is 4 times that of Pfu DNA polymerase, with better fidelity and higher heat resistance temperature. It is the most efficient thermostable DNA polymerase. KOD DNA polymerase has 3'-5' exonuclease activity, that is, proofreading function, which ensures that it can quickly identify and correct incorrectly incorporated nucleotides. This product has no 5'-3' exonuclease activity.
Features
Excellent fidelity
Actual sequencing results show that in the PCR process of KOD DNA Polymerase, the frequency of base mismatches is lower, and the fidelity of KOD DNA Polymerase is about 20 times higher than that of Taq DNA Polymerase or other companies' Long PCR enzymes.
Excellent elongation (possible chain length for amplification)
40kb can be amplified using λDNA as template, 24kb can be amplified using human genomic DNA as template, and 12kb can be amplified using cDNA as template.
Superior amplification efficiency
Compared with previous PCR enzymes, the yield is greatly improved, so amplification can be performed even with a small amount of template.
High reliability (PCR success rate)
Target fragments with high GC content, whole blood, etc. can be directly amplified, and the PCR success rate is very high.
Transportation and storage
This product is shipped with dry ice and can be stored at -15℃~ -25℃ for two years.
PCR experimental steps
(1) Preparation of PCR reaction solution
Before preparing the reaction solution, please fully mix all reagents except KOD DNA Polymerase. Please use frozen reagents after they are completely thawed.
（2）Reaction system (50 µL/reaction)
|
Components |
Final concentration |
Dosage |
|
5×PCR Buffer |
1 × |
10 µL |
|
dNTP Mix |
200 µM |
1 µL |
|
Upstream primer |
0.2 µM |
1 µL |
|
Downstream primer |
0.2 µM |
1 µL |
|
DNA template |
- |
x µL |
|
KOD DNA Polymerase |
0.04U/µL |
0.4 µL |
|
sterile water |
- |
Up to 50 µL |
Recommended template usage
|
Template type |
Recommended dosage |
|
Genomic DNA |
10 ng-200 ng |
|
Plasmid or viral DNA |
10 pg-50 ng |
|
cDNA |
1-5 μL (no more than 1/10 of the total PCR volume) |
（3）Reaction procedure
①Three-step PCR
*Set the annealing temperature according to the primer TM value. If the primer TM value is ≥72℃, the annealing step can be deleted and the subsequent extension step (two-step PCR) can be directly performed. If necessary, a temperature gradient can be established to find the optimal temperature for the primer to bind to the template. In addition, the annealing temperature directly determines the amplification specificity. If the amplification specificity is found to be poor, the annealing temperature can be appropriately increased.
*For most templates, 30s/kb can be effectively amplified. For some complex templates, the extension time can be increased to 30~60s/kb
②Two-step PCR
Precautions
1. Please do not use primers and templates containing uracil.
2. In order to improve the success rate and yield of amplification, please use high-quality templates.
3. It can be dissolved at room temperature. After dissolution, it is advisable to store it in an ice box or on an ice bath. After use, it is advisable to store it at -20ºC immediately.
4. For your safety and health, please wear a lab coat and disposable gloves when operating.
| Components | GE10004-01 | GE10004-02 | GE10004-03 | GE10004-04 |
| KOD DNA Polymerase | 50 μL | 100 μL | 200 μL | 500 μL |
| 5 × PCR Buffer | 500 μL | 500 μL | 1 mL | 1 mL*3 |
Quality Control & SDS
- View current batch:
-
Purity: >95.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
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