FPCRN-RO

FastStart^™ Taq DNA Polymerase, dNTPack

• Synonym(s): DNA Polymerase, DTNPs, PCR

Properties

• usage: sufficient for ≤1,250 reactions (04738403001); sufficient for ≤2,500 reactions (04738420001); sufficient for ≤250 reactions (04738357001); sufficient for ≤50 reactions (04738314001); sufficient for ≤500 reactions (04738381001)
• Quality Level: 100
• packaging: pkg of 1,000 U (04738381001 [4x250 U]); pkg of 2,500 U (04738403001 [10x250 U]); pkg of 5,000 U (04738420001 [20x250 U]); pkg of 100 U (04738314001); pkg of 500 U (04738357001 [2x250 U])
• manufacturer/tradename: Roche
• parameter: 72 °C optimum reaction temp.
• storage temp.: −20°C

Description

General description

The enzyme dNTPack comprises FastStart Taq DNA Polymerase and a ready-to-use solution of PCR grade nucleotides. FastStart Taq DNA Polymerase is a versatile enzyme that can be used in a wide variety of applications and on multiple instrument platforms. This modified, thermostable, recombinant Taq DNA polymerase is inactive at temperatures below +75 °C, but is activated by a 2 to 4 minute heat activation step at +95 °C. Since it is inactive at low temperatures, FastStart Taq DNA Polymerase cannot elongate non-specific primer-template hybrids that may form at those temperatures. An optimized PCR buffer system and a GC-RICH Solution to enable the enzyme to handle a wide range of templates are also supplied.

Application

FastStart^™ Taq DNA Polymerase is a thermostable, chemically modified form of recombinant Taq DNA Polymerase. The enzyme is inactive at +15 to +25°C during PCR setup, and then activated at +95°C during initial denaturation. This enzyme delivers superior results due to its unique enzyme design and optimized buffer system. FastStart Taq DNA Polymerase is an ideal tool for hot start PCR, because the enzyme remains inactive during PCR setup and prior to the initial denaturation step. It can be applied for PCR:

• Hot start PCR up to 3 kb
• Hot Start RT-PCR up to 3 kb
• Multiplex PCR
• Difficult templates, e.g., secondary structures or GC-rich sequences
• Automated PCR, e.g., handling at room temperatures
• quantitaive PCR(qPCR)

For maximum convenience, select the 2x concentrated ready-to-use FastStart PCR Master.

Features and Benefits

Volume activity: 5 U/μl
Each dNTPack contains 10 mM additive-free sodium salt nucleotides as a ready-to-use mix.

• Higher specificity, sensitivity, and yield:

Hot start PCR makes PCR setup easier.

• Use robotic setup.

Use this enyzme mix stable for 24 hours at +15 to +25°C.

• Prevent PCR carryover contamination.

Incorporate dUTP and use Uracil-DNA Glycosylase to pretreat PCR master mixes.

• Cost-effective.

Use the convenient premixed solution of PCR grade nucleotides.

Packaging

1 kit containing 6 components

Quality

Each lot is function-tested using human genomic DNA and primers specific for the 365 bp fragment of human tPA gene, and, a 284 bp fragment of the Apo E gene with 74% GC content. Each lot is also tested for the absence of exo- and endonucleases and nicking activity.
For details please refer to the respective Instruction for Use.

Unit Definition

One unit FastStart DNA Polymerase is defined as the amount of enzyme that incorporates 10 nmol of total deoxyribonucleosidetriphosphates into acid precipitable DNA within 30 minutes at +75 °C under the assay conditions stated under unit assay.

Unit Assay: 1 μg M13mp9ss DNA, 0.3 μg M13 sequencing primer and 0.1 μCi [α--^32P] dCTP are incubated with varying amounts of units of FastStart Taq DNA Polymerase in 50 μl incubation buffer at 65 °C for 60 min. The amount of incorporated dNTPs is determined.

Volume Activity: 5 U/μl

Other Notes

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

Hot start protocols improve PCR specificity, sensitivity, and yield. Heat-labile blocking groups make the modified enzyme inactive at +15 to +25°C. No elongation occurs when primers bind nonspecifically. The enzyme is activated by removing the blocking groups at +95°C for 2 to 6 minutes. PCR products with 3′-single A overhangs are produced, ideal for T/A cloning. For PCR carryover prevention, use the PCR Nucleotide Mix^PLUS and Uracil-DNA Glycosidase, heat-labile.

Legal Information

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

Kit Components Only

• FastStart Taq DNA Polymerase, in storage and dilution buffer 5 U/μl
• PCR Reaction Buffer, with 20 mM MgCl₂ 10x concentrated
• PCR Reaction Buffer, without MgCl₂ 10x concentrated
• MgCl₂ Stock Solution 25 mM
• GC-RICH Solution 5x concentrated
• PCR Nucleotide Mix

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): does not flash
• Flash Point(C): does not flash