PCR Applications

GC-Rich PCR

Break the Bond

When do you need it?

GC-rich PCR concerns DNA templates with high GC content, which is defined as the percentage of guanine-cytosine base pairs. When amplifying templates with >60% GC content (i.e 5' ends of most mammalian cDNAs), the three hydrogen bonds shared by GC pairs lead to enhanced thermostability, which is problematic for un-optimized Taq polymerase systems.

How is it achieved?

Destabilizing GC base pairs is the key to tackling GC-rich PCR. This is achieved by supplementing the reaction buffer with GC-melt and DMSO to disrupt or destabilize base pairing, and by using a robust polymerase mix that maintains processivity in the presence of these additives.

What makes our PCR polymerases the best?

We offer high performance enzyme mixes and kits for GC-rich PCR, suited for templates with up to 90% GC content.

Advantage GC 2 Kit includes the Advantage 2 Polymerase Mix, our proprietary GC-Melt reagent, and reaction buffer containing DMSO. These components work in concert to amplify highly complex GC-rich targets with minimal buffer optimization.
Advantage GC Genomic LA Mix is an enzyme blend of full-length, thermostable Taq DNA Polymerase, a small amount of proofreading enzyme includes an additional GC-Melt buffer system, designed specifically to amplify GC-rich templates over 6 kb.

How to pick the right enzyme?
Polymerase	Application	Fidelity*	Amplicon size (kb) Human/plasmid/cDNA	Max GC content	Hot Start	Cloning compatibility
Advantage GC 2	GC-rich PCR	3X	<6.0/<6.0/<2.0	90%	Yes	TA
Advantage GC Genomic LA	Long, GC-rich PCR	6.5X	<8.0–30.0/<20.0/<10.0	90%	Yes	TA
* Relative fidelities are represented as fold improvements over Titanium Taq DNA Polymerase, which has an error rate of 1 mismatch per kb of synthesized DNA.