Sigma-Aldrich Faststart Hifi PCR System

Usage

sufficient for ≤200 reactions (04738292001)
sufficient for ≤50 reactions (04738284001)

Quality Level

100

Packaging

pkg of 125 U (04738284001)
pkg of 500 U (04738292001 [2 x 250 U])

Manufacturer/Tradename

Roche

Parameter

72 °C optimum reaction temp.

Technique(S)

PCR: suitable

Storage Temp.

−20°C

Related Categories

PCR Reagents & Kits

Roche® Life Science Products

General Description

The enzyme dNTPack comprises the FastStart High Fidelity PCR System and a ready-to-use solution of PCR grade nucleotides. The FastStart High Fidelity PCR System is a blend of FastStart Taq DNA Polymerase and a thermostable proofreading protein, which is also chemically modified. This protein mediates proofreading activity, but carries no polymerase activity. Both proteins are inactive below +75°C and are activated by heating to +95°C for two minutes. The FastStart^®High Fidelity PCR System is the ideal tool for all amplification experiments where a combination of high specificity, sensitivity, accuracy, and yield is needed. Hot start protocols have improved specificity, sensitivity, and PCR yield. Heat-labile blocking groups on amino acid residues of FastStart Taq DNA Polymerase make the enzyme inactive at room temperature. There is no elongation at low temperatures when primers bind nonspecifically.

Application

The FastStart^®High Fidelity PCR System – an optimized enzyme blend for hot start PCR – combines all the benefits of FastStart Taq DNA Polymerase with fourfold higher accuracy and the ability to amplify fragments up to 5kb.

• Hot start PCR
• Multiplex PCR
• Hot Start RT-PCR
• GC-rich template amplification
• Difficult or challenging PCRs
• precapture ligation-mediated PCR (LM-PCR)
• PCR amplification of cDNA 16S rRNA gene[5][6]

The FastStart High Fidelity PCR System is the product of choice for multiplex PCR[7]

Features And Benefits

• Achieve excellent multiplex performance: For fast high quality results, also use Roche′s PCR Optimization Kit.
• Increase fidelity: Achieve fourfold higher fidelity compared to Taq DNA Polymerase and FastStart Taq DNA Polymerase.
• Amplify challenging DNA: DMSO, a PCR additive, for working with difficult templates, is supplied.
• Cost-effective: Use the convenient premixed solution of PCR grade dNTPs.

Packaging

1 kit containing 6 components

• FastStart High Fidelity Enzyme Blend, (5 U/μl) in storage buffer
• FastStart High Fidelity Reaction Buffer, 10x concentrated, with 18mM MgCl₂
• FastStart High Fidelity Reaction Buffer, 10x concentrated, without MgCl₂
• MgCl₂Stock Solution, 25mM
• DMSO
• PCR Nucleotide Mix (each dNTPack contains the 10mM additive-free sodium salt nucleotides in a ready-to-use mix)

Quality

Each lot is PCR function-tested using human genomic DNA and primers specific for a 1.8 kb fragment of the EPO gene, and primers specific for a 4.8 kb fragment of the tPA gene.

Unit Definition

Volume Activity: 5 U/μl

Preparation Note

Working concentration:For a standard 50 μl PCR, we recommend using 2.5 U of the enzyme blend.

Other Notes

Contents

• FastStart High Fidelity Enzyme Blend, (5 U/μl) in storage buffer
• FastStart High Fidelity Reaction Buffer, 10x concentrated, with 18 mm MgCl₂
• FastStart High Fidelity Reaction Buffer, 10x concentrated, without MgCl₂
• MgCl₂Stock Solution, 25 mm
• DMSO
• PCR Nucleotide Mix

Each dNTPack contains the 10 mm additive-free sodium salt nucleotides in a ready-to-use mix.NOTICE TO PURCHASER: LIMITED LICENSE<br />Use of this product is covered by US Patent No. 6,127,155 and corresponding patent claims outside the US. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Human and veterinary diagnostic uses under Roche patent claims require a separate license from Roche. All uses other than internal research and human and veterinary diagnostic uses under Roche patent claims require a separate license from Thermo Fisher Scientific. Further information on purchasing licenses from Roche may be obtained by contacting the Licensing Department of Roche Molecular Systems, Inc., 4300 Hacienda Drive, Pleasanton, California 94588, USA or Roche Diagnostics GmbH, Sandhofer Strasse 116, 68305 Mannheim, Germany. Further information on purchasing licenses from Thermo Fisher Scientific may be obtained by contacting the Licensing Department of Thermo Fisher Scientific, 5791 Van Allen Way, Carlsbad, California 92008, USA.</ br>NOTICE TO PURCHASER: LIMITED LICENSE<br />Use of this product is covered by US Patent Nos. 5,677,152 and 5,773,258, and corresponding patent claims outside the US. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Human and veterinary diagnostic uses under Roche patent claims require a separate license from Roche. All uses other than internal research and human and veterinary diagnostic uses under Roche patent claims require a separate license from Thermo Fisher Scientific. Further information on purchasing licenses from Roche may be obtained by contacting the Licensing Department of Roche Molecular Systems, Inc., 4300 Hacienda Drive, Pleasanton, California 94588, USA or Roche Diagnostics GmbH, Sandhofer Strasse 116, 68305 Mannheim, Germany. Further information on purchasing licenses from Thermo Fisher Scientific may be obtained by contacting the Licensing Department of Thermo Fisher Scientific, 5791 Van Allen Way, Carlsbad, California 92008, USA.FastStart is a trademark of RocheRoche is a registered trademark of Roche Molecular Systems, Inc.