SKU : G3444-200U
หมวดหมู่ : Biochemicals ,  1. Chemical and Reagents ,  Buffers ,  Servicebio , 
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Product Information
Product Name | Cat. No. | Spec. |
Fast High Fidelity DNA Polymerase | G3444-200U | 200 U |
Product 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 Shipping Conditions
Ship with wet ice; Store at-20 up to 12 months.
Product Contents
Component Number | Component | G3444-200U |
G3444-1 | Fast High Fidelity DNA polymerase | 100 μL |
G3444-2 | 10х Reaction buffer | 1 mL |
G3444-3 | dNTP(10 mM each) | 500 μL |
Manual | One copy | |
Assay Protocol/Procedures
Commonly used PCR reaction system (50 μL) :
Component | Volume |
10х Reaction buffer | 5 μL |
Nuclease-Free water | To 50 μL |
dNTP Mix (10 mM each) | 1 μL |
Forward primer/Reverse primer (10 uM) | 2 μL/2 μL |
Template | n ( Adjustable ) |
Fast High Fidelity DNA polymerase | 0.5 μL ( Adjustable ) |
Note: For 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. The 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. for high GC template no more than 10 % DMSO can be added to the system. Other volume of reaction system can be adjusted according to the 50μL reaction system.
Recommended PCR procedures :
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 | 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.
For Research Use Only!
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