FastStart Universal Probe Master (Rox)

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:

• 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

Combine this master mix with Transcriptor First Strand cDNA Synthesis Kit (Roche) to achieve excellent results in two-step qRT-PCR.

• Increase qPCR sensitivity and specificity.

Produce lower cycle threshold (Ct) values.

• 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.

Use a master mix that is stable at room temperature during extended reaction setup times.

• 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.

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).

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.

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.