G-HiFi DNA Polymerase, High GC High Fidelity DNA Polymerase

High GC High Fidelity DNA Polymerase, High GC DNA Template DNA Polymerase (Cat# TF3000). 1U/ul, 100U/Pack

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

• Long range PCR amplification for next-generation DNA sequencing
• Generates blunt end amplicons for cloning with GetClone™ PCR cloning vector
• Amplification of GC-rich templates

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

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.