The UCP Multiplex PCR Kit provides a convenient ready-to-use master mix formulation and UCP PCR water. It also contains orange UCP Master Mix Tracer and blue UCP Template Tracer in separate vials for optional use. (see figure
''UCP Multiplex PCR Kit''). The master mix includes a balanced combination of K+ and NH4+ ions as well as the unique synthetic Factor MP, which together promote stable and efficient annealing of primers to the nucleic acid template, enabling high PCR efficiency of all targets (see figure
"Unique PCR buffer"). Successful separation of 20 targets or more can be achieved by traditional gel or capillary electrophoresis, while sequencing-based detection strategies can accommodate significantly higher multiplexing degrees. At low temperatures, the DNA Polymerase is kept in an inactive state by an antibody and a novel Guard additive, which stabilizes the complex. (see figure
''Principle of Guard-protected hot-start mechanism''). This improves the stringency of the hot-start and prevents any enzymatic activity at ambient temperature, allowing reaction setup without ice or on instruments. The enzyme is fully activated after the 2-minute incubation step at 95°C and starts amplifying with high specificity from the first cycle. The hot-start procedure prevents extension from non-specifically annealed primers and primer-dimers in the first cycle ensuring highly specific and reproducible PCR. The blue and orange dyes in the UCP Template Tracer and in the UCP Master Mix Tracer, respectively, allow visual tracking of pipetted samples during the PCR setup, to prevent errors. When the blue template is added to the orange PCR Master Mix, the color changes to green, confirming that sample was added. (see figure
''Visual pipetting control''). The use of these tracers is optional. Reactions can be directly loaded onto agarose gels after cycling. Each tracer dye allows monitoring of the loading process and efficient tracking of the subsequent electrophoresis.
