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).
Universal ready-to-use hot start reaction mix for qPCR and RT-qPCR on all real-time PCR systems requiring normalization with ROX.
FastStart™ Universal Probe Master (Rox) includes a novel reference dye that enables its use on all real-time PCR instruments requiring normalization with ROX, without modification or adjustments to the specific instrument or protocol. 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. This product is not intended for use with the LightCycler® Instruments.
FastStart™ Universal Probe Master (Rox) has been used:
Combine this master mix with Transcriptor First Strand cDNA Synthesis Kit (Roche) to achieve excellent results in two-step qRT-PCR.
- in TaqMan® quantitative real-time polymerase chain reaction (qRT-PCR) reactions for the quantification of endogenous miRNAs, such as MIR376B, MIR376A and MIR181A
- in reverse transcriptase (RT-PCR) to study tumor necrosis factor (TNF) expression in whole synovial tissue of undifferentiated peripheral inflammatory arthritis (UPIA) patients
- for the amplification and detection of any DNA or cDNA target, including those that are GC- or AT-rich by quantitativePCR
Features and Benefits
Produce lower cycle threshold (Ct) values.
- Increase qPCR sensitivity and specificity.
Achieve sensitive, specific results in assays with the Universal ProbeLibrary Probes or any other hydrolysis probe.
- Use the master mix with any probe-based assay.
Amplify fragments up to 500 bp long, including those that are GC- or AT-rich.
- Amplify and detect a broad range of DNA or cDNA targets.
- Visualize amplification products on agarose gels.
Use a master mix that is stable at room temperature during extended reaction setup times.
- Use robotic pipetting stations to set up qPCR reactions.
Use this dUTP-containing mix with Uracil-DNA Glycosylase to eliminate contaminating DNA carried over from previous PCR reactions.
- Prevent false positives resulting from carryover contamination.
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.
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).
For life science research only. Not for use in diagnostic procedures
FastStart is a trademark of Roche
LightCycler is a registered trademark of Roche
TaqMan is a registered trademark of Roche Molecular Systems, Inc.