GC-RICH PCR System

for amplification of GC-rich targets (up to 5 kb) from genomic DNA Cat. No. 2 140 306 50 PCR reactions

Description

Unique enzyme blend of Taq DNA Polymerase and Pwo DNA Polymerase[a thermostable enzyme with a proofreading (3'–5' exonuclease) activity].
The GC-RICH PCR System also includes:

An optimized GC-RICH PCR reaction buffer, 5x conc. containing 7.5 mM MgCl₂ and DMSO
GC-RICH resolution solution, 5 M
MgCl₂ stock solution, 25 mM
PCR grade water

Application

The GC-RICH PCR System is designed to amplify difficult DNA templates up to 5 kb including:

GC-rich targets
Repetitive sequences
Templates with varying GC content (multiplex PCR, construction of random libraries)

Note: The GC-RICH PCR System may also be used in standard PCR, where it will give better results (higher yield and greater fidelity) than Taq DNA Polymerase alone.

Application profile

Operating parameters

Starting sample: 10–500 ng DNA or 1–100 ng cDNA
Temperature optimum (for elongation):
- 72°C (for targets < 3 kb)
- 68°C (for targets > 3 kb)
GC-RICH PCR System accepts modified nucleotides like DIG-dUTP

Experimental result

Successfully amplify high GC-rich templates with the GC-RICH PCR System. Amplification of a 264 bp template (GC content 74%) within the human ApoE gene using the GC-RICH PCR System or the Expand High Fidelity PCR System.

Key advantages

Allows amplification of difficult templates like GC-rich or repetitive sequences up to 5 kb, because the enzyme blend and the buffer provided with the system are optimized for these templates
Ideal for multiplex PCR, because the GC-RICH PCR System amplifies all targets with similar efficiency, regardless of GC content
Improves fidelity of PCR amplification, because the GC-RICH PCR System transcribes with threefold greater accuracy than Taq DNA Polymerase alone

Roche Contact Information

Roche

Roche Diagnostics Corporation
P.O. Box 50414
9115 Hague Road
Indianapolis, IN 46250-0414 USA

Customer Service: 800-262-1640

Tech Support: 800-262-4911

Fax Number: 800-428-2883

Web Site: http://www.roche-applied-science.com

GC-RICH PCR System