High-Fidelity PCR Kits

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PCR Master Mixes and Kits

Overview

iProof™ Polymerase Benefits

Comparison of common thermostable DNA polymerases. iProof Polymerase exhibits the highest fidelity of any commercially available thermostable polymerase, >50-fold higher than ordinary Taq polymerase. A common high-fidelity polymerase was ~8-fold higher than Taq, and an ultrahigh-fidelity polymerase was ~32-fold higher.

iProof Polymerase demonstrates unrivaled speed, leading to dramatically shorter overall reaction times. The reaction protocol for iProof Polymerase was compared to the recommended protocols for two competing polymerases. Each protocol was designed to amplify 1, 8, or 15 kb products in 30 cycles. Reactions with iProof Polymerase used a two-step protocol with a combined annealing and extension step, while the other reactions used three-step protocols with the minimum recommended extension times. Overall reaction times include temperature ramping times.

iProof DNA Polymerase Performance Data

Comparison of amplification using iProof Polymerase and another supplier's fast- cycling enzyme. Manufacturer's recommended cycling conditions were used for both enzymes. Left, a 1 kb fragment was amplified from 10 ng human genomic DNA using 20 U/ml of iProof Polymerase (lanes 6–9) or 50 U/ml of the other enzyme (lanes 20–24) in a standard-ramping thermal cycler. Total run time for protocols using a 30 sec annealing/extension step was 48 min (lanes 6, 7, 20, and 21). Total run time for protocols using a 15 sec annealing/extension step was 39 min (lanes 8 and 9) or 40 min (lanes 22 and 23). Lane 5, 500 bp EZ Load marker. Right, the same 1 kb fragment was amplified from 100 ng human genomic DNA on a fast-ramping thermal cycler, using 50 U/ml of iProof Polymerase (lane 3) or 100 U/ml of the other enzyme (lane 4), and using an annealing/extension step of 30 sec. Total run time on the fast ramping cycler was 30 min. Lane 2, 500 bp EZ Load Marker.

iProof DNA polymerase amplifies long templates with high yields.
Left, various fragments up to 37 kb in length were amplified from BAC DNA using a combined annealing/extension step of 10 min per cycle and 30 U/ml of iProof polymerase. Right, various sequences up to 28.8 kb were amplified directly from human genomic DNA using 30 U/ml of iProof polymerase in GC buffer with a combined annealing/extension time of 10 min per cycle.

Annealing/extension time, sec
iProof High-Fidelity Polymerase Competitor High-Fidelity Polymerase Taq Polymerase

iProof DNA Polymerase delivers high yields with significantly less enzyme. A 1 kb fragment was amplified from 1.25 pM of lambda DNA template in 20 cycles, using combined annealing/extension step of variable duration as indicated. All enzymes were used according to manufacturer recommendations.

High-fidelity PCR reagents featuring the thermostable iProof Polymerase. This enzyme utilizes Sso7d fusion polymerase technology to provide highly accurate amplification of targets up to 37 kb.


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Description

iProof High-Fidelity DNA Polymerase is a unique Pyrococcus-like proofreading enzyme fused to the Sso7d dsDNA-binding protein to create a thermostable fusion polymerase that accurately amplifies long products from a variety of DNA templates.

Features and Benefits of iProof High-Fidelity DNA Polymerase:

  • High fidelity — iProof DNA polymerase is 52-fold more accurate than Taq polymerase
  • Speed — high processivity dramatically reduces extension (15–30 sec/kb) and overall reaction times
  • Successful amplification of long products with higher yields — fragments up to 37 kb are amplified in less time and with less enzyme (0.25–1 unit/reaction)

Choose the best DNA polymerase for your application.

Key Features iProof DNA Polymerase iTaq DNA Polymerase
Speed

Processivity

Long fragment amplification

Proofreading Yes No
Hot-Start No Yes
 

Related Categories

Supporting Documentation


Specifications

Storage at –20°C iProof™ Enzyme, Kit, and Buffers: guaranteed for 6 months at –20°C in a constant-temperature freezer
iProof Master Mixes: guaranteed for 3 months at –20°C in a constant-temperature freezer (multiple freeze/thaws not recommended)
Storage at +4°C Guaranteed for 3 months at 4°C

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iProof HF Master Mix, 100 x 50 µl rxns, 2.5 ml

1725310
100 x 50 µl reactions, premixed PCR reagents, includes 2x master mix (0.04 U/µl), DMSO; suitable for most templates

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$225.00 
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iProof GC Master Mix, 100 x 50 µl rxns, 2.5 ml

1725320
100 x 50 µl reactions, premixed PCR reagents, includes 2x master mix (0.04 U/µl), DMSO; suitable for GC-rich templates

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$332.00 
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iProof™ High-Fidelity DNA Polymerase, 20 U (2 U/µl), 10 µl

1725300
20 U, 2 U/µl, stand-alone enzyme, includes 5x reaction buffers, MgCl2 solution, DMSO

This product is being discontinued soon. Please contact customer service for more information.

The following replacement is recommended:

  • iProof™ High-Fidelity DNA Polymerase, 100 U (2 U/µl), 50 µl (1725301)
iProof™ High-Fidelity DNA Polymerase, 100 U (2 U/µl), 50 µl

1725301
100 U, 2 U/µl, stand-alone enzyme, includes 5x reaction buffers, MgCl2 solution, DMSO

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$146.00 
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iProof™ High-Fidelity DNA Polymerase, 500 U (2 U/µl), 250 µl

1725302
500 U, 2 U/µl, stand-alone enzyme, includes 5x reaction buffers, MgCl2 solution, DMSO

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$579.00 
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iProof™ High-Fidelity PCR Kit, 50 U (2 U/µl), 25 µl

1725330
50 U, 2 U/µl, iProof polymerase PCR kit includes 5x reaction buffers, MgCl2 solution, DMSO, dNTPs, λ DNA, 1.3 and 10 kb primers, DNA standard

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$85.00 
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iProof™ High-Fidelity PCR Kit, 200 U (2 U/µl), 100 µl

1725331
200 U, 2 U/µl, iProof DNA polymerase PCR kit includes 5x reaction buffers, MgCl2 solution, DMSO, dNTPs, λ DNA, 1.3 and 10 kb primers, DNA standard

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$309.00 
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Documents

Number Description Options
10002300 Instruction Manual, iProof High-Fidelity PCR Kit, Rev B Click to download
10002298 Instruction Manual, iProof High-Fidelity DNA Polymerase, Rev B Click to download
5211 iProof High-Fidelity DNA Polymerase Flier, Rev C Click to download [ Add to Cart (Free) ]
6252 Reagent Comparison Guide for Real-Time PCR Brochure, Rev C Click to download [ Add to Cart (Free) ]
6090 Amplification Reagents and Plastics Brochure Click to download [ Add to Cart (Free) ]
5337 Long Inverse PCR Using iProof Polymerase, Rev A Click to download
10000083948 Instruction Manual, iProof™ High-Fidelity PCR Master Mix Click to download

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