High-throughput PCR: EmeraldAmp GT PCR Master Mix

EmeraldAmp GT PCR Master Mix is a loading-dye-added version of EmeraldAmp MAX PCR Master Mix that is optimized for great performance and convenience in both standard and high-throughput PCR applications. This master mix includes an optimized buffer, PCR enzyme, dNTP mixture, gel loading dye (green), and a density reagent in a 2X premix format. Simply add primers and DNA template. Following PCR, amplicons can be used directly in several ways.

PCR tube contents can be loaded directly onto an agarose gel for electrophoresis or used directly in downstream applications such as restriction enzyme digestion, TA cloning, and direct sequencing. EmeraldAmp GT PCR Master Mix can be used to amplify genomic targets up to ~5 kb and is compatible with GC- and AT-rich targets.

When 5 µl of the loading-dye-added PCR master mix is used for electrophoresis on an 1% Agarose L03 gel, the blue dye front is detected around 3–5 kb, and the yellow dye is detected below 50 bp. These dyes have absorptions at approximately 260 nm and 420 nm, respectively. The dyes can be removed by gel purification or PCR cleanup using the NucleoSpin Gel and PCR Clean-Up kit (where available), if necessary.

Overview

• Convenient premix format—just add template, primers, and water to the 2X PCR master mix
• Improved performance—includes an optimized buffer for better amplification of GC- and AT-rich targets
• Eliminates purification steps—PCR contents can be loaded directly onto an agarose gel
• High-throughput PCR—use reactions directly in TA cloning, restriction enzyme digestion, or sequencing
• Green loading dye—allows tracking during gel electrophoresis

Applications

High-throughput PCR

PCR products

Most PCR products amplified with EmeraldAmp GT have a 3'-terminal adenosine (A), and therefore PCR products can be used directly for cloning into T-vectors (TA cloning). It is possible to clone the product into blunt-end vectors after blunting and phosphorylation.