BioMaster HS-qPCR Hi-ROX SYBR (2×) contains 2× BioMaster HS-qPCR Hi-ROX SYBR reaction mix, 50 mM MgCl2 solution and sterile water. The mix is developed for quantitative real-time PCR with the use of fluorescent dye SYBR Green I on PCR platforms that utilize ROX as a reference dye. BioMaster HS-qPCR Hi-ROX SYBR (2×) includes all the components (except for DNA template and primers) necessary for PCR: highly processive recombinant HS-Taq DNA polymerase, dNTP mix, PCR buffer, Mg2+ solution, SYBR Green I, fluorescent dye ROX, and inert dye.
BioMaster HS-qPCR Hi-ROX SYBR (2×) is optimized for efficient and reproducible hot-start real-time PCR of genomic, plasmid and viral DNA samples. BioMaster HS-qPCR Hi-ROX SYBR (2×) includes additives that influence primer annealing temperature and characteristics of template melting thus enabling to increase PCR specificity and use templates with complicated spatial structure.
DNA polymerase included in the mix is inactive at room temperature and its activation requires preheating at 95 °C for 5 min. Blue hue of the reaction solution provided by the inert dye allows control when using multi-well plates. Use of the mix saves time and minimizes contamination risk due to reduced number of pipetting steps. Low concentration of Mg2+ in the reaction solution and additional tube with 50 mM MgCl2 solution allow optimization of reaction conditions for individual primer/template system.
The master mix is ideally suitable for PCR platforms that use ROX passive dye as a reference guide, such as Life Technologies (ABI) 7000, 7300, 7700, 7900, 7900HT, and StepOnePlus.