- SG/ROX onTaq qPCR Master Mix (2x) is a universal solution for quantitative real-time PCR and two-step real-time RT-PCR and can be used on most real-time PCR cyclers available regardless of requirements in ROX reference dye.
- The master mix contains onTaq DNA Polymerase, optimized reaction buffer, dNTPs (dTTP is partially replaced with dUTP), SYBR Green I dye and ROX passive reference dye.
- SYBR Green I enables DNA analysis without having to synthesize expensive sequence-specific labeled probes.
- onTaq DNA Polymerase is a modified „hot start” enzyme that is blocked at moderate temperatures and allows room temperature reaction setup. The polymerase activity is restored during 15 min initial denaturation step at 95°C.
- Use of the “hot start” enzyme prevents extension of misprimed products and primer-dimers during reaction setup leading to higher specificity and sensitivity of PCR reactions.
- The polymerase enables convenient room temperature reaction setup.
- SYBR Green I is a fluorescent dye which binds all doublestranded DNA molecules and emits a fluorescent signal on binding. The excitation and emission maxima of SYBR Green I are at 494 nm and 521 nm, respectively, which are compatible with use on any real-time cycler.
- SG/ROX onTaq qPCR Master Mix (2x) contains dUTP, which partially replaces dTTP. It allows the optional use of a uracil-N -glycosylase (UNG) to prevent carryover contamination between reactions. UNG removes uracilfrom any dU-containing contaminating amplicons, leaving abasic sites and making DNA molecules susceptible to hydrolysis during the initial denaturation step.
- ROX passive reference dye included in the master mix allows fluorescence normalization on certain cyclers. The use of ROX dye is necessary for all real-time PCR cyclers from Applied Biosystems and optional for cyclers from Agilent. ROX compensates for variations of fluorescent signal between wells due to slight differences in reaction volume and
fluorescence fluctuations. ROX is not involved in PCR reaction, has a different emission spectrum than SYBR Green I and does not interfere with real-time PCR on any instrument.
- SG/ROX onTaq qPCR Master Mix (2x) is intended to use with most cyclers from Applied Biosystems, Bio-Rad, Cepheid, Roche, Eppendorf, Agilent, Qiagen, etc.