Sigma-Aldrich Faststart Universal Probe

Usage

sufficient for 1250 reactions
sufficient for 250 reactions
sufficient for 5000 reactions

Quality Level

100

Feature

dNTPs included: no
hotstart

Manufacturer/Tradename

Roche

Packaging

pkg of 1250 x 20 μL reactions (04913957001)
pkg of 250 x 20 μL reactions (04913949001)
pkg of 5000 x 20 μL reactions (04914058001)

Technique(S)

RT-qPCR: suitable
qPCR: suitable

Input

purified DNA

Detection Method

probe-based

Related Categories

PCR Reagents & Kits

Roche® Life Science Products

General Description

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.

Application

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[5][6]

Combine this master mix with Transcriptor First Strand cDNA Synthesis Kit (Roche) to achieve excellent results in two-step qRT-PCR.[7]

Features And Benefits

• 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.

Components

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).

Other Notes

For life science research only. Not for use in diagnostic proceduresFastStart is a trademark of RocheLightCycler is a registered trademark of RocheTaqMan is a registered trademark of Roche Molecular Systems, Inc.