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Maxima® SYBR Green qPCR Buffer has been specifically optimized for qPCR analysis using SYBR® Green I. It contains both KCl and (NH4)2SO4 to provide high specificity of primer annealing. The buffer composition allows for PCR at a wide range of MgCl2 concentrations. Therefore, optimization of MgCl2 concentration in PCR is generally not necessary.
SYBR® Green I is a fluorescent intercalating dye which binds to the double stranded DNA and emits a fluorescent signal upon binding. In qPCR, DNA accumulates and fluorescent signal increases proportionally to the DNA concentration. The excitation and emission maxima of SYBR® Green I are at 494 nm and 521 nm, respectively, which are compatible with the use on any real-time cycler.
dUTP is included in the master mix to partially replace dTTP in the accumulated PCR product, allowing for the option to prevent carryover contamination between reactions (1). Uracil-DNA Glycosylase (UDG) pre-treatment of the reaction removes all dU-containing amplicons carried over from previous reactions.
Note. UDG is not included in the Maxima® SYBR Green qPCR Master Mix, no ROX, and must be purchased |