ALLin™ RPH Polymerase, 5 u/µl

Applications


  • Long PCR (up to 35 kb) with higher fidelity

  • Amplification of complex (GC rich) templates

  • Crude sample PCR, multiplex PCR, TA cloning

Benefits


  • Long (up to 35 kb) high-fidelity (5X higher than Taq) amplification

  • High yields in standard and fast PCR, on GC rich DNA and crude samples

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

Catalog. # Product Name Size Price Qty
HLE0101 ALLin™ RPH Polymerase, 5 u/µl 250 u
€199.50
HLE0105 ALLin™ RPH Polymerase, 5 u/µl 1250 u
€899.50

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Description

highQu ALLin™ RPH Polymerase (Robust, Proofreading, Hot-start Polymerase) is the versatile engineered enzyme combining best polymerase properties for excellence in most demanding PCR applications, like low copy detection, long or high fidelity PCR, amplification of complex templates, crude sample PCR and multiplexing.

ALLin™ RPH Polymerase has 5 times higher fidelity than Taq DNA Polymerase and produces A-tailed products suitable for ligating into TA cloning vectors.

For the maximum convenience the ALLin™ RPH Mastermix, 2X is available.

 

 

 

 

Protocols

Print version of the protocol: Product Insert ALLin™ RPH Polymerase, 5 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: 15 sec/kb - <5kb and 40 to 60 sec/kb - <5kb-35 kb. Use >90 sec extension for multiplexing.
  • Run an annealing temperature gradient from 55°C to 65°C to choose the best specificity conditions.
  • Do not use fast cycling for multiplexing.

 

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™ RPH Buffer 10 µl
Water to 49 μl
ALLin™ RPH Polymerase, 5 u/µl 0.25 - 1 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 – ~10 min (for 10 kb)

15 sec/kb for <5 kb amplicons

40-60 sec/kb for >5 kb amplicons

 

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

Specifications

CAT.

SIZE

COMPONENTS

COMPOSITION

HLE0101 250 u 250 u - ALLin™ RPH Polymerase, 5 u/µl
2 x 1 ml - 5X ALLin™ RPH Buffer
Enzyme in storage buffer.
1X ALLin™ RPH Buffer contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers.
HLE0105 1250 u 5 x 250 u - ALLin™ RPH Polymerase, 5 u/µl
10 x 1 ml - 5X ALLin™ RPH Buffer
Enzyme in storage buffer.
1X ALLin™ RPH Buffer contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers.


Storage


In the dark at -20°C.

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