DNA Polymerase High Fidelity (HiFi) Enzyme

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Catalogue No: abx071010
Price: US$304.50
(Size: 250 U)

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Datasheet SDS
DNA Polymerase High Fidelity (HiFi DNA Polymerase) contains T-DNA Polymerase and a proofreading 3'-5' exonuclease. This kit contains two different buffers: HiFi Buffer 1 is optimized for the amplification of genomic DNA, and HiFi Buffer 2 is optimized for the amplification of λDNA, cDNA or plasmid DNA. The extension rate is about 1-2 kb/min. Template-independent "A" can be generated at the 3' end of the PCR product. PCR products can be directly cloned into T-vectors. Genomic DNA fragments can be amplified up to 15 kb. This kit does not contain 2.5 mM dNTPs.

Contents:

Component 250 U 500 U 3 kU
HiFi DNA Polymerase250 U 500 U 6 × 500 U
10X HiFi Buffer 1 1.2 ml 1.2 ml 6 × 1.2 ml
10X HiFi Buffer 2 1.2 ml 1.2 ml 6 × 1.2 ml
10X GC Enhancer 200 µl 400 µl 1 ml
6X DNA Loading Buffer 500 µl 1 ml 2 × 1 ml


Target DNA Polymerase (HiFi)
Tested Applications PCR
Conjugation Unconjugated
Purity > 99% (SDS-PAGE)
Quality Control Assayed for amplication efficiency to amplify the p53 gene from 10 ng of human genomic DNA.
Storage Store at -20 °C for up to 2 years. Avoid repeated freeze/thaw cycles.
Buffer HiFi DNA Polymerase: 20 mM Tris-HCl (pH 8.0), 0.1 mM EDTA, 1 mM DTT, 100 mM KCl, 50% glycerol, stabilizers.
10X HiFi Buffer 1/2: 200 mM Tris-HCl (pH 9.0), 100 mM (NH4)2SO4, 20 mM MgSO4, 100 mM KCl, 10% glycerol, other proprietary ingredients.
Biological Activity One unit of HiFi DNA Polymerase incorporates 10 nmol of deoxyribonucleotide into acid-precipitable material in 30 minutes at 74 °C.
Endotoxin Level Functional absence of double and single stranded endonuclease activity.
Concentration 5 U/µl
Availability Shipped within 10-20 working days.
Note This product is for research use only.

Not for human consumption, cosmetic, therapeutic or diagnostic use.
Directions for use Reaction Components:

Component Volume Final Concentration
Template Variable as required
Forward Primer (10 µM) 1 µl 0.2 µM
Reverse Primer (10 µM) 1 µl 0.2 µM
10X HiFi Buffer 1/2 5 µl 1X
2.5 mM dNTPs 4 µl 0.2 mM
HiFi DNA Polymerase 0.5-1 µl 2.5-5 U
Nuclease-free H2O Variable N/A
Total Volume 50 µl N/A

Thermal Cycling Conditions:

Number of Cycles Temperature Time
1 cycle 94 °C 2-5 min
30-35 cycles 94 °C 30 seconds
50-60 °C 30 seconds
72 °C 1-2 kb/min
1 cycle 72 °C5-10 min

Notes:
  • For GC/AT-rich or complex templates, it is recommended to add GC Enhancer to the PCR reaction mixture. The suggested working concentration range for the 10X GC Enhancer provided in this kit is 0.5-5X.
  • A final concentration of 2 mM MgSO4 is sufficient to amplify most targets. Some targets may require a higher concentration of Mg2+
  • For optimal results, we recommend using a 100 mM MgSO4 stock solution to prepare a titration from 2 mM to 4 mM (final concentration) in 0.25 mM increments.
  • 0.5 µl (2.5 U) of enzyme is sufficient for a reaction vlume of 50 µl. For increased amplification, up to 1 µl (5 U) of enzyme can be used.
Research Articles on DNA Polymerase (HiFi)


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