Sigma-Aldrich Faststart High Fidelity Pc

Usage

sufficient for ≤1,000 reactions (03553361001)
sufficient for ≤200 reactions (03553400001)

Quality Level

100

Feature

High Fidelity PCR
Multiplex PCR
dNTPs included: no
hotstart

Packaging

pkg of 2,500 U (03553361001 [10 x 250 U])
pkg of 500 U (03553400001 [2 x 250 U])

Manufacturer/Tradename

Roche

Concentration

5 unit/μL

Parameter

72 °C optimum reaction temp.

Technique(S)

PCR: suitable

Input

purified DNA

Storage Temp.

−20°C

Related Categories

PCR Reagents & Kits

Roche® Life Science Products

General Description

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.

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. FastStart^™Taq DNA Polymerase is activated by removal of blocking groups using a preincubation step at +95°C. The FastStart^™High Fidelity PCR System is the product of choice for multiplex PCR.

• 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

Application

The FastStart^™High Fidelity PCR System — a new enzyme blend for hot start PCR — combines all the benefits of FastStart^™Taq DNA Polymerase with fourfold higher accuracy and the abilty to amplify fragments up to 5 kb.

• PCR
• Multiplex PCR
• RT-PCR
• GC-rich template amplification
• Difficult or challenging PCRs
• in the amplification of Human Papillomaviral genome[5] and methylated DNA[6]

The FastStart^™High Fidelity PCR System is the product of choice for multiplex PCR.
Use FastStart^™High Fidelity PCR System, dNTPack with ready-to-use PCR Nucleotide Mix.

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.

Packaging

1 kit containing 5 components

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

One unit of FastStart Taq DNA Polymerase is defined as the amount of enzyme that incorporates 10 nmol of total desoxyribonucleotide triphosphates into acid precipitable DNA within 60 min at +65 °C

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

For life science research only. Not for use in diagnostic procedures.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 Roche