Application

FastStart^™ Universal SYBR^® Green Master (Rox) can be used for the amplification and detection of any DNA or cDNA target, including those that are GC- or AT-rich. Combine this master mix with Transcriptor First Strand cDNA Synthesis Kit (Roche) to achieve excellent results in two-step qRT-PCR.FastStart^™ Universal SYBR^® Green Master (Rox) has been used in qRT-PCR and qPCR

Components

FastStart Universal SYBR Green Master (Rox), 2x concentrated master mix that contains FastStart Taq DNA Polymerase, Reaction Buffer, Nucleotides (dATP, dCTP, dGTP, dUTP), SYBR Green I, and a reference dye.

Features and Benefits

Improve PCR sensitivity and specificity.Minimize the formation of nonspecific amplification products. Avoid over-estimation of qPCR results.Eliminate nonspecific amplification products and primer-dimers that increase the amount of bound quantified SYBR Green I. Amplify and detect a broad range of DNA or cDNA targets.Amplify fragments up to 500 bp long, including those that are GC- or AT-rich. Save time and effort in qPCR preparation.Eliminate the need to mix components, titrate MgCl₂, or perform other time-consuming optimization steps. Prevent false positives resulting from carryover contamination.Use this dUTP-containing mix with Uracil-DNA Glycosylase to eliminate contaminating DNA carried over from previous PCR reactions.

General description

FastStart^™ Universal SYBR^® Green Master (Rox) is a ready-to-use hot start reaction mix for qPCR and RT-qPCR on all real-time PCR systems requiring normalization with ROX. SYBR^® Green I is a DNA double-strand-specific dye. During each phase of DNA synthesis, the SYBR^® Green I dye, which is included in the reaction mix, binds to the amplified PCR products. The amplicon can be detected by its fluorescence. Hot start protocols have been shown to significantly improve the specificity, sensitivity, and yield of PCR. Heat-labile blocking groups on some of the amino acid residues of FastStart^™ Taq DNA Polymerase make the modified enzyme inactive at room temperature (+15 to +25°C). Therefore, there is no elongation during the period when primers can non-specifically bind. FastStart^™ Taq DNA Polymerase is activated by removing the blocking groups at a high temperature (i.e., a pre-incubation step at +95°C).

Legal Information

SYBR is a registered trademark of Life Technologies

FastStart is a trademark of Roche

Other Notes

For life science research only. Not for use in diagnostic procedures.

Quality

Function test: Each lot is tested for performance in qPCR using three templates: a GC-rich template, an AT-rich template, and a long template (approximately 440 bp).