The High GC High Fidelity DNA Polymerase, G-HiFi™ DNA Polymerase is a new genetically modified, recombinant DNA polymerase suitable for GC-rich templates that are difficult to amplify. The fidelity of G-HiFi™ DNA Polymerase is 70 times higher than that of Taq DNA polymerase. The high extension rate of G-HiFi™ DNA Polymerase is achieved by blending the DNA polymerase with an elongation enhancer. The optimized 5X G-HiFi™ Buffer includes special ingredients that suppress non-specific amplification as well as plateau effect produced by conventional PCR. With the optimized 5X G-HiFi™ Buffer, G-HiFi™ DNA Polymerase is capable to amplify most templates, such as longer targets (up to 40 kb from lambda DNA) and that contain GC-rich sequences.
Features
• 5’→3’ DNA polymerase activity
• 3’→5’ exonuclease (proofreading) activity
• Suitable for GC-rich templates
• High reaction rate: 7 seconds/kb
• High fidelity: 70 times higher than Taq polymerase
• Generates blunt end amplicons
• Vast elongation capability (up to 40 kb)
• Thermo-stable for more than 10 hrs at 95°C.
Fig. 1. G-HiFiTM DNA Polymerase’s high processability enables reliable amplification of λDNA up to 40 kb in length (M: DM5100).
Fig. 2. G-HiFiTM DNA Polymerase performs higher sensitivity for high GC content templates (GC: 71%) compare to high fidelity DNA Polymerase from Brand A (M: DM2000).
Applications
Pack Contents
100 ul Q-HiFi High Fidelity DNA Polymerase, 1U/ul
1200 ul 5× Q-HiFi Reaction Buffer
600 ul dNTPs Mix (2mM each)
Storage Buffer
50 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizers, and 50% (v/ v) glycerol
Unit Definition
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 74°C.
Storage
-20°C ≥ 24 months
Recommended PCR Conditions
Template |
1 ~ 150 ng |
Forward primer |
0.1 ~ 0.5 μM |
Reverse primer |
0.1 ~ 0.5 μM |
5 × Reaction Buffer |
10 μl |
dNTPs |
0.2 mM (each) |
Q-HiFi High Fidelity DNA Plolymerase |
1 μl |
H2O |
to 50 μl |
Total volume |
50 μl |
Recommended PCR Program For ≦10 kb products:
98°C 2 min
98°C 15 sec
50~68°C* 30 sec
68°C 10 ~30 sec/kb
68°C 1 min
*Optimal PCR condition varies according to primers’ thermodynamic properties.
Recommended PCR Program For ≧10 kb products:
98°C 10 sec
68°C 10 ~30 sec/kb
Recommended Primer design
For ≦10 kb products:
For general amplification, select primers with a Tm value of ≧ 55°C. 20- to 25-mer primers are suitable, or greater than 25-mer in length may provide optimal results.
For >10 kb products:
Select primers with a Tm value of ≧ 65°C. 25- to 35-mer primers are suitable. Avoid high GC-content at the 3′ end of each primer.
Shipping cost
Please note this item is NOT participating the “Free Shipping on $200” rule. A $40 invoice will be sent for a standard 2nd-day shipping. If you order our other Taq PCR related products we only charge shipping cost once. Check our other Taq PCR related products. Buy more save more.
Reviews
There are no reviews yet.