An efficient way to determine the presence or absence of a target sequence—often needed for genotype screening—is by visualizing end-point PCR products using agarose gel electrophoresis and ethidium bromide staining. When screening for dozens (or even hundreds) of samples, efficiency is critical, and every improvement in the workflow leads to a significant saving in time. When setting up a large number of PCR reactions, eliminating pipetting steps reduces the potential for contamination and error and minimizes repetitive strain-induced injury. Simplifying the screening procedure also enables better standardization between users, which can be crucial as screening projects scale up.
Takara Bio's PCR premixes provide the flexibility and convenience required for high sample throughput by minimizing pipetting steps. EmeraldAmp GT PCR Master Mix is suitable for amplifying GC- or AT-rich genomic DNA products up to 5 kb in length. For amplifying targets longer than 5 kb, EmeraldAmp MAX PCR Master Mix can be used, which is optimized for long targets and includes a loading dye.
EmeraldAmp GT PCR Master Mix provides excellent performance at an affordable price. PCR reactions are easy to assemble by simply adding primers and template to the 2X premix which contains a buffer, PCR polymerase, dNTPs, gel loading dyes, and a density reagent. After PCR, samples can be loaded directly onto an agarose gel for electrophoresis, used for restriction digestion, or easily cloned into a vector.
We compared the performance of Takara Bio's EmeraldAmp premixes to another dye-added premix Taq on the market, Bioline's MyTaq Red Mix. We looked at parameters such as amplification of long targets (up to 4 kb), GC-rich targets, and nonspecific amplification. The results are presented below.