ALLin™ HiFi DNA Polymerase, 2 u/µl

Applications


  • High-fidelity PCR up to 10 kb

  • Complex (GC rich) template or crude sample PCR

  • Fast PCR

  • Blunt cloning

Benefits


  • 50 x higher fidelity than classic Taq

  • High sensitivity and yields under standard and fast cycling

  • Increased success in long (10 kb) or GC rich PCR

  • 5X ALLin™ Buffer contains optimal Mg2+ and dNTPs

Catalog. # Product Name Size Price Qty
HLE0201 ALLin™HiFi DNA Polymerase, 2 u/µl 200 u
€149.50
HLE0205 ALLin™HiFi DNA Polymerase, 2 u/µl 1000 u
€672.50

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Description

highQu ALLin™ HiFi DNA Polymerase is the outperforming high-fidelity enzyme derived from Pfu polymerase by introducing several point mutations. The robust engineered enzyme in combination with the optimized ALLin™ buffer provides higher fidelity (50X higher than Taq DNA Polymerase), better performance, increased success in demanding applications like amplification of complex or longer templates, crude samples and fast cycling.

ALLin™ HiFi DNA Polymerase produces blunt-ended products suitable for ligating into blunt vectors.

Protocols

Print version of the protocol: Product Insert ALLin™HiFi DNA Polymerase, 2 u/µl
Important Notes
  • Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips.
  • Include a no-template control and positive control in parallel.
  • Thaw and keep reagents on ice. Mix well before use.
  • The longer the amplicon, the longer the extension time: 30 sec/kb.
  • Run an annealing temperature gradient from 55°C to 65°C to choose the best specificity conditions.

 

Prepare a 50 µl PCR reaction


Rev. & For. Primers variable, up to 0.4 µM final each (2 µl of 10 µM each)
cDNA Template  or
gDNA Template
<100 ng  or
5 - 500 ng
5X ALLin™ HiFi Buffer 10 µl
Water to 49 μl
ALLin™ HiFi DNA  Polymerase, 2 u/µl 0.5 µl

 

  • Mix gently, avoid bubbles.
  • Place into the instrument set like:

 

Initial denaturation 1 cycle: 95°C – 1 min
Denaturation 25-35 cycles: 95°C - 15 sec
Annealing 25-35 cycles: 55-65°C – 15 sec
Extension 25-35 cycles: 72°C –30 sec (30 sec/kb)

 

  •  After the PCR store probes shortly on ice, for long term store at -20°C.

Specifications

CAT. SIZE COMPONENTS COMPOSITION
HLE0201 200 u 200 u - ALLin™ HiFi DNA Polymerase, 2 u/µl
3 x 1 ml - 5X ALLin™ HiFi Buffer
Enzyme in storage buffer.
1X ALLin™ HiFi Buffer contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers.
HLE0205 1000 u 5 x 200 u - ALLin™ HiFi DNA Polymerase, 2 u/µl
15 x 1 ml - 5X ALLin™ HiFi Buffer
Enzyme in storage buffer.
1X ALLin™ HiFi Buffer contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers.


Storage


In the dark at -20°C.

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