GC RICH PCR SYSTEM

General description

The GC-RICH PCR System is composed of an enzyme blend of thermostable Taq DNA Polymerase and Tgo DNA Polymerase, a thermostable enzyme with a proofreading (3′-5′ exonuclease) activity. This polymerase mixture by itself outperforms Taq DNA Polymerase in respect to yields, fidelity and specificity beside the possibility to amplify fragments up to 5 kb in length. The GC-RICH PCR Reaction buffer in combination with the separately included GC-RICH resolution solution allows to amplify difficult templates like GC-rich targets very efficiently. TA cloning is recommended. The enzyme blend produces more blunt-ended fragments than Taq DNA Polymerase. The majority of products have single A overhangs. Use 2 U for a standard 50 μl PCR.

Contents

• GC-RICH Enzyme Mix , in storage buffer
• GC-RICH Reaction Buffer, 5x concentrated, with 7.5 mM MgCl2 and DMSO
• GC-RICH Resolution Solution, 5 M
• MgCl2 Stock Solution, 25 mM
• Water, PCR Grade

Application

GC-RICH PCR system has been used in semi‐quantitative analysis, genotyping and retrieval and cloning of high-GC content sequence of candidate gene.

The GC-RICH PCR System, a blend of Taq DNA polymerase and a proofreading polymerase, enables amplification of templates that are difficult or impossible to amplify with other polymerases and other blends of polymerases. The enhanced processivity of the blend and the unique GC-RICH Resolution Solution combine to deliver superb performance – especially from problematic templates.
The GC-RICH PCR System may also be used in standard PCR applications, providing improved results (higher yield, higher accuracy) over Taq DNA polymerase alone.

Advantages of the GC-Rich PCR System:

• Ease access to difficult templates, including GC-rich targets and repetitive sequences.
• Reagents, the GC-RICH Resolution Solution and PCR Grade Water are provided.
• Amplify DNA fragments up to 5 kb.

Features and Benefits

Blend of Taq DNA Polymerase and a proofreading polymerase amplifies difficult templates. Enhanced processivity and the unique GC-RICH Resolution Solution deliver superior performance.

• PCR
• Difficult templates up to 5 kb

The GC-RICH PCR System is a blend of Taq DNA Polymerase and a proofreading polymerase for amplifying longer nucleic acid fragments. The GC-Rich Solution provides an solution to enable amplification of all kind of difficult PCR products.
When using the supplied magnesium-containing buffer, the final MgCl2 concentration is 1.5 mM.

Packaging

1 kit containing 5 components

Quality

The GC-RICH PCR System is function-tested, by amplifying a human 284 bp ApoE genomic fragment, using the GC-RICH PCR System protocols

Unit Definition

Volume Activity: 2 U/μl

Preparation Note

Working concentration: Enzyme concentration
The optimal enzyme concentration range from 0.5 to 5 U per assay. For a standard 50 μl PCR, we recommend using 2 U of the enzyme blend.

Other Notes

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

Legal Information

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.