10X GC-Rich PCR Enhancer Buffer

Cat No.: DCP-GCEB10X

Size: 20 mL
Sterility: Non-Sterile, 0.22-micron membrane filtered once
Storage: -20ºC (Aliquot into smaller volumes for long-term storage at −20°C)
Shelf Life: 1 year

• For working stocks at 4°C, shelf life is 6 months.
• Avoid repeated freeze-thaw cycles to maintain stability.

pH: 8.3-8.7
Shipping: Dry Ice

The 10X Enhancer Buffer is a specialized PCR additive that improves amplification of high GC-content DNA templates by destabilizing secondary structures and equalizing GC/AT melting behavior, often via betaine formulation. Use at 1X–3X final concentration in reactions; calibrate optimal levels per template via testing.

• Best to use the reagent and perform Gradient PCR to find the optimal temperature at which the reagent works the best.
• Best to perform serial dilutions to find the optimal template-to-primer ratio for high GC-rich regions. Suggested dilutions are 0, 1:5, 1:25, 1:100, and 1:500 dilutions of the template DNA. GC-rich regions with PCR enhancers typically work in this dilution range.  
• Other optimizations may be required for a DNA region rich in GC, as well as forming strong secondary structures.
• Strongly suggest Hot-Start or similar PCR reagents (DiagnoCine offers many top-notch PCR reagents) [HERE])

Key Benefits:

• Lowers GC-rich melting temperatures to match AT-rich regions for better primer binding and polymerase progression.
• Boosts specificity and yield for challenging templates.
• Minimizes DNA polymerase pausing in GC tracts.
• Reduces secondary structure formation for greater template accessibility.

Usage:

1.  Use the GC?rich PCR Enhancer at 10×, 7.5×, 5×, and 2.5× in parallel reactions to systematically determine which enhancer concentration gives the strongest and cleanest amplification.
2.  Perform a gradient PCR in which you vary both the annealing temperature and the enhancer level (10×, 7.5×, 5×, 2.5×) to find the best conditions for each primer pair on the GC?rich template.
3.  From these experiments, identify the specific combination of annealing temperature (Tm) and enhancer concentration that provides the most efficient, specific amplification for the given GC?rich DNA target.

Usage: Add 1 µL per 10 µL reaction


* This product is manufactured under ISO 13485-certified and CE-approved facilities (Suppliers of DiagnoCine Precision). All final packaging, quality assurance, and testing are done at the DiagnoCine R&D and Quality Testing Center.  All specific customization requests and assembly were accomplished at DiagnoCine Precision in Totowa, New Jersey, USA