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Product Description
The FastStart Universal SYBR Green Master (Rox) is a ready-to-use reagent mix that simplifies the preparation of reactions for qPCR and two-step qRT-PCR using the intercalating dye SYBR Green I for DNA detection and analysis. In combination with a real-time PCR instrument and suitable PCR primers, FastStart Universal SYBR Green Master (Rox) enables very sensitive detection and quantification of defined DNA sequences.
This product is not intended for use with the LightCycler®Instruments.
In principle, the 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-rich or AT-rich.
Combine this master mix with our Transcriptor First Strand cDNA Synthesis Kit to achieve excellent results in two-step qRT-PCR. Insist on the Transcriptor First Strand cDNA Synthesis Kit – designed and function-tested for qPCR, and efficient with all real-time PCR instruments.
Application
FastStart Universal SYBR Green Master (Rox) includes a novel reference dye, which enables its use without modifications or adjustments to the specific instrument or protocol on all real-time PCR instruments requiring normalization with ROX. This ready-to-use, 2x-concentrated master mix contains all reagents (except primers and template) needed for running quantitative, real-time DNA detection assays, including qPCR and two-step qRT-PCR, in the SYBR Green I detection format. FastStart Universal SYBR Green Master (Rox) generates excellent results on instruments such as the Applied Biosystems 7900 HT Fast Real-Time PCR System or the Applied Biosystems 7500 Real-Time PCR System.
Background Information
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. Therefore, there is no elongation during the period when primers can nonspecifically bind. The FastStart Taq DNA Polymerase is activated by removing the blocking groups at a high temperature (i.e.,a pre-incubation step at 95°C).