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ALLin™ RPH Mastermix

Kód produktu: HLM0101 Kód výrobce: HLM0101 Kód dodavatele: {641C524E-2236-439D-9C06-3A3CC383338C} Výrobce: highQu GmbH professionally simple
-4 %
Ušetříte
336,38 Kč
9 082,26 Kč
8 745,88 Kč
7 228,00 Kč bez DPH
do týdne
200

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.

Zobraz detailní popis
  • Zeptejte se odborníka

Detailní popis

Applications


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

  • Amplification of complex (GC rich) templates

  • Crude sample PCR, multiplex PCR, TA cloning



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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

  • ALLin™ mastermix supplied with PCR Water

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.

The convenience of ALLin™ RPH Polymerase is maximized by the use of 2X Mastermix providing the additional advantage of reduced pipetting and minimized errors. The mastermix is even supplied with PCR Water, and the only thing to add is the template with primers.

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
PCR Water to 25 μl
ALLin™ RPH Mastermix, 2X 25 µl

 

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

 

Initial denaturation 1 cycle: 95°C - 1-2 min
Denaturation 25-40 cycles: 95°C - 15 sec
Annealing 25-40 cycles: 55-65°C- 15 sec
Extension

25-40 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.
CAT. SIZE COMPONENTS COMPOSITION
HLM0101 200 r
of 50 µl
5 x 1 ml - ALLin™ RPH Mastermix, 2X
5 x 1 ml - PCR Water
1X mastermix contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers.
HLM0105 1000 r
of 50 µl
25 x 1 ml - ALLin™ RPH Mastermix, 2X
25 x 1 ml - PCR Water
1X mastermix contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers.

 

Storage

 

In the dark at -20°C.

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