Fast High Fidelity DNA polymerase
Cat. No.: SB-G3444-200U
Size: 200U
Description
Fast High Fidelity DNA polymerase is a engineered ultra-high fidelity DNA polymerase combining with single-stranded DNA-binding protein. It possesses the fast extension speed available up to 5-15 s/kb, along with the extremely high fidelity 50-fold higher than of Taq, and 6-fold higher than that of Pfu. with 5'-3' polymerase and 3'-5' exonuclease activities, it produces biunt end DNA products. It is ideal for routine PCR, colony PCR, Amplification of difficult (GC-rich) templates and Long-range amplification up to 14 kb.
- Definition of activity unit: The amount of enzyme required to incorporate 10 nmol of deoxynucleotide into acid insoluble material is defined as one unit of enzyme activity at 74°C for 30 min using activated salmon sperm DNA as a template primer.
- Purity and concentration: ≥ 95% detected by SDS-PAGE; endogenous nucleic acid < 1 pg/μL detected by qPCR.
- Concentration：2 U/μL.
Storage and transportation
Transport in wet ice packs; store at -20°C, valid for 12 months.
Product Contents
| Component Number |
Component |
G3444-200U |
| G3444-1 |
Fast High Fidelity DNA polymerase |
100 μL |
| G3444-2 |
10× Reaction buffer |
1mL |
| G3444-3 |
dNTP（10 mM each） |
500 μL |
| Product Manual |
1 copy |
Assay Protocol/Procedures
Commonly used PCR reaction system (50 μL):
| Component |
Volume |
| template |
n (adjustable) |
| Fast High Fidelity DNA polymerase |
0.5 μL (adjustable) |
| dNTP Mix (10 mM each) |
1 μL |
| Forward primer/Reverse primer (10 μM) |
2 μL/2 μL |
| 10× Reaction buffer |
5 μL |
| Nuclease Free Water |
To 50 μL |
Note: ①For plasmid or phage DNA as a template, the recommended addition amount is 10 ng-1 pg per 50 μL system; For genomic DNA as a template, the recommended addition amount is 250 ng-50 ng per 50 μL system; For cDNA as a template, it is recommended to dilute 2-100 folds, the addition amount does not exceed 10 % of the final volume ; For bacterial liquid or crude extract sample as a template, add no more than 10 % of the reaction volume. Excessive templates are easy to lead to non-specific amplification, too few templates are easy to lead to low efficiency of PCR amplification. ②The final concentration range of primers is 0.2-1.0μM, and the recommended primer concentration is 0.4 μM. Too few primers may lead to low or no amplification, and excessive primers may lead to non-specific amplification. ③for high GC template no more than 10 % DMSO can be added to the system. ④Other volume of reaction system can be adjusted according to the 50μL reaction system.
Recommended PCR reaction procedure:
| Temperature |
Time |
Cycles |
| 98℃ |
3 min |
1 |
| 98℃ |
15-30 s |
25 - 35 |
| 50℃ - 72℃ |
15-30 s |
| 72℃ |
5-15 s/kb |
| 72℃ |
5-10 min |
1 |
| 4-16°C |
Forever |
|
Note
1. Enzyme products should be kept in an ice box or ice bath when used, and stored at -20℃ immediately after use. It is recommended to store them separately.
2. For your safety and health, please wear a lab coat and disposable gloves.
