Clontech Laboratories, Inc. • www.clontech.com
Reprinted from Clontechniques October 2006
CR6X2171 US (630604)
Advantage® GC Genomic LA Polymerase Mix
Long and accurate genomic DNA amplification for GC-rich templates
• Efficiently and accurately amplifies
GC-rich regions
• Optimized buffer system
with Advantage GC-Melt
• Fast extension rate
It is often difficult to amplify long, highly
GC-rich templates by standard methods.
Clontech’s Advantage GC Genomic LA
Polymerase Mix combines the benefits
of our Advantage Genomic LA Polymerase
mix, such as high efficiency and improved
fidelity on long templates, with our
GC-Melt reagent’s ability to amplify
GC-rich sequences.
Highly Efficient for GC-Rich
Templates
GC-rich templates have strong second-
ary structures which resist denaturation
and prevent efficient primer annealing.
Clontech’s Advantage GC-Melt buffer
system weakens base pairing in GC-rich
sequences (Figure 1). This allows efficient
amplification of virtually all GC-rich
sequences (up to 90% GC) that resist
standard PCR amplification techniques.
The Advantage Genomic LA Polymerase
enzyme blend was used with two different
buffers to amplify three highly GC-rich
sequences: c-jun, 65% GC; TGF-β, 69%
GC; and IGFR2, containing a 100 bp
region that is 90% GC (Figure 2). In the
presence of the Advantage GC-Melt buf-
fer, the expected fragments were ampli-
fied from human genomic DNA (c-jun
and TGF-β), or from a cDNA template
(IGFR2), with high efficiency (Lanes 1,
3, and 5). Amplification was much less ef-
ficient with standard Advantage Genomic
LA Buffer (Lanes 2, 4, and 6). As shown
in Figure 2, the Advantage GC-Melt
Buffer provided with this mix can be the
difference between success or failure when
amplifying GC-rich templates.
Highly Accurate & Processive
Amplification
The Advantage GC Genomic LA Poly-
merase Mix includes a full length, thermo-
stable Taq polymerase and a small amount
of DNA polymerase with 3' to 5' proof-
reading activity. The proofreading enzyme
removes mis