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FastStart Universal Probe Master (Rox)
Universal ready-to-use hot start reaction mix for qPCR and qRT-PCR on all real-time PCR systems requiring normalization with ROX.
Background Information
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. FastStart Taq DNA Polymerase is activated by removing the blocking groups at a high temperature (i.e., a preincubation step at 95°C).
Contents
FastStart Universal Probe Master (Rox), 2x-concentrated master mix that contains FastStart Taq DNA Polymerase, Reaction Buffer, Nucleotides (dATP, dCTP, dGTP, dUTP), and a reference dye.
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).
Application
FastStart Universal Probe Master (Rox) includes a novel reference dye, which enables its use without modification 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, probe, and template) needed for running quantitative, real-time DNA-detection assays, including qPCR and two-step qRT-PCR, in the hydrolysis probe detection format. FastStart Universal Probe 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.
Benefits
•Use this master mix on all real-time PCR instruments that require normalization:
Benefit from the new, specially developed universal reference dye included in the master mix for robust performance with a variety of instrument platforms.
•Increase qPCR sensitivity and specificity:
Produce lower cycle threshold (Ct) values with the hot start enzyme FastStart Taq DNA Polymerase and highest purity nucleotides included in the master mix.
•Use the master mix with any probe-based assay:
Achieve sensitive, specific results in assays with the Universal ProbeLibrary Probes or any other hydrolysis probe.
•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.
•Visualize amplification products on agarose gels.
•Use robotic pipetting stations to set up qPCR reactions:
Benefit from a master mix that is stable at room temperature during extended reaction setup times.
•Prevent false positives resulting from carryover contamination:
The mix contains dUTP, so that it may be used with Uracil-DNA Glycosylase to eliminate contaminating DNA carried over from previous PCR reactions. |