The UltraRun LongRange PCR Kit provides a convenient 4x master mix formulation, an orange Master UCP Mix Tracer, a blue UCP Template Tracer and Q-Solution (for optional use) in separate vials.
The ready-to-use master mix consists of a blend of enzymes – Taq DNA polymerase and a polymerase with proofreading capability – which improves hands-on time and process robustness, a sophisticated PCR buffer optimized for long PCR and dNTP mix. At low temperatures, both polymerase and proofreader activities are kept in an inactive state by antibodies. This provides a stringent hot-start and prevents any enzymatic activity until heat activation at 93°C, thereby avoiding primer or DNA damage from untimely proofreader activity. Due to this functionality, complete reactions, including primers and template, are stable at room temperature for longer periods for convenient process handling. Furthermore, the 4x master mix concentration allows greater flexibility for template input volumes over common 2x master mixes and the dyes in the master mix formulation enable direct loading of PCR reactions onto agarose gels for analysis.
The UltraRun LongRange PCR Kit includes Q-Solution, an innovative PCR additive that facilitates amplification of difficult templates by modifying the melting behavior of DNA. Unlike other commonly used PCR additives such as DMSO, Q-Solution is used at just one working concentration, is non-toxic and PCR purity is guaranteed.
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 the 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.