iProof™ High-Fidelity DNA Polymerase

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Overview

Specifications
iProof™ Enzyme, Kit, and Buffers
Storage at –20°C Guaranteed for 6 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
iProof Master Mixes
Storage at –20°C 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
Exceptional Speed

Comparison of amplification using iProof 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 (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 (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.

Higher Fidelity Available

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

Long Fragment Amplification

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. Right, various sequences up to 28.8 kb were amplified directly from human genomic DNA using 30 U/ml of iProof in GC buffer with a combined annealing/extension time of 10 min per cycle.

Higher Yields — Less Enzyme

iProof polymerase demonstrates increased speed with minimal amounts of enzyme. The gel images below compare the performance of three thermostable polymerases in amplifying a 1 kb DNA fragment using a two-step protocol that combined annealing and extension times ranging from 7 to 240 sec. iProof polymerase successfully amplified the target with the shortest extension time (15 sec). Competing enzymes required an annealing/extension step of 30–60 sec or more and 5- to 10-fold more enzyme to produce comparable yields.

Annealing/extension time, sec
iProof high-fidelity polymerase Common 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.

Unrivaled Speed

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 high-fidelity DNA polymerase comprises a unique Pyrococcus-like proofreading enzyme fused to a dsDNA binding protein, Sso7d. This results in a thermostable polymerase that accurately amplifies long products from a variety of DNA templates. iProof polymerase is available in three convenient formats: a stand-alone enzyme, an easy-to-use master mix, and a PCR kit complete with controls. In comparison to reactions using Taq polymerase, all three formats feature:

  • High fidelity — iProof DNA polymerase is 52-fold more accurate
  • 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)
iProof™ High-Fidelity DNA Polymerase

172-5300
20 U, 2 U/µl, stand-alone enzyme, includes 5x reaction buffers, MgCl2 solution, DMSO

List Price:   $30.00
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iProof™ High-Fidelity DNA Polymerase

172-5301
100 U, 2 U/µl, stand-alone enzyme, includes 5x reaction buffers, MgCl2 solution, DMSO

List Price:   $139.00
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iProof™ High-Fidelity DNA Polymerase

172-5302
500 U, 2 U/µl, stand-alone enzyme, includes 5x reaction buffers, MgCl2 solution, DMSO

List Price:   $598.00
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iProof™ HF Master Mix

172-5310
100 x 50 µl reactions, premixed PCR reagents, includes 2x master mix (0.04 U/µl), DMSO; suitable for most templates

List Price:   $226.00
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iProof™ GC Master Mix

172-5320
100 x 50 µl reactions, premixed PCR reagents, includes 2x master mix (0.04 U/µl), DMSO; suitable for GC-rich templates

List Price:   $306.00
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iProof™ High-Fidelity PCR Kit

172-5330
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

List Price:   $90.00
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iProof™ High-Fidelity PCR Kit

172-5331
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

List Price:   $312.00
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5x iProof HF Buffer

172-5391
1.5 ml, 5x iProof high-fidelity PCR buffer

List Price:   $29.00
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5x iProof GC Buffer

172-5392
1.5 ml, 5x iProof PCR buffer for GC-rich templates

List Price:   $29.00
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5x iProof HPLC HF Buffer

172-5393
1.5 ml, detergent-free for HPLC, 5x iProof high-fidelity PCR buffer

List Price:   $29.00
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5x iProof HPLC GC Buffer

172-5394
1.5 ml, detergent-free for HPLC, 5x iProof PCR buffer for GC-rich templates

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Number Description Options
6252 Reagent Comparison Guide for Real-Time PCR Brochure, Rev C Click to download
6090 Amplification Reagents and Plastics Brochure, Rev D Click to download [ Add to Cart (Free) ]
5337 Long Inverse PCR Using iProof Polymerase, Rev A Click to download
5211 iProof High-Fidelity DNA Polymerase Flier, Rev C Click to download [ Add to Cart (Free) ]
10002298 Instruction Manual, iProof High-Fidelity DNA Polymerase, Rev B Click to download
10002299 Instruction Manual, iProof High-Fidelity Master Mix, Rev E Click to download
10002300 Instruction Manual, iProof High-Fidelity PCR Kit, Rev B Click to download
5856 Mutational Analysis of the ATM Gene in Familial Breast Cancer Using iProof High-Fidelity DNA Polymerase, Rev A Click to download [ Add to Cart (Free) ]