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FastStart Universal Probe Master (Rox)

sufficient for 250 reactions, sufficient for 1250 reactions, sufficient for 5000 reactions, suitable for qPCR, suitable for RT-qPCR

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About This Item

UNSPSC Code:
41106300
NACRES:
NA.55

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

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

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 procedures

Legal Information

FastStart is a trademark of Roche
LightCycler is a registered trademark of Roche
TaqMan is a registered trademark of Roche Molecular Systems, Inc.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Stefano Alivernini et al.
Frontiers in medicine, 5, 186-186 (2018-07-19)
Objectives: To examine synovial tissue (ST) predictors of clinical differentiation in patients with seronegative undifferentiated peripheral inflammatory arthritis (UPIA). Methods: Fourty-two patients with IgA/IgM-Rheumatoid Factor and anti-citrullinated peptide antibodies negative UPIA, naive to Disease-Modifying Anti-Rheumatic Drugs, underwent Gray Scale (GSUS)
The essential function for serum response factor in T-cell development reflects its specific coupling to extracellular signal-regulated kinase signaling.
Mylona A, et al.
Molecular and Cellular Biology, 31(2), 267-276 (2011)
Matthew L Hillestad et al.
Human gene therapy, 23(10), 1116-1126 (2012-07-28)
Reporter genes are important tools for assessing vector pharmacology in vivo. Although useful, current systems are limited by (1) the need to generate a new vector for each different reporter, (2) the inability to package reporter genes in small vectors
Robert J Ihry et al.
Nature medicine, 24(7), 939-946 (2018-06-13)
CRISPR/Cas9 has revolutionized our ability to engineer genomes and conduct genome-wide screens in human cells1-3. Whereas some cell types are amenable to genome engineering, genomes of human pluripotent stem cells (hPSCs) have been difficult to engineer, with reduced efficiencies relative

Articles

Hot Start PCR

The purpose of Hot Start PCR is to inhibit the PCR reaction in order to reduce nonspecific amplification, prevent the formation of primer dimers, and increase product yields.

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PCR Applications

Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.

RT-qPCR – Quantitative Reverse Transcription PCR

RT-qPCR, or quantitative reverse transcription PCR, combines the effects of reverse transcription and quantitative PCR or real-time PCR to amplify and detect specific targets. RT-qPCR has a variety of applications including quantifying gene expression levels, validating RNA interference (RNAi), and detecting pathogens such as viruses.

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