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L6669

Sigma-Aldrich

LuminoCt® qPCR ReadyMix™

For fast probe-based quantitative PCR

Sinónimos:

LuminoCt® qPCR ReadyMix™, LuminoCt®

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About This Item

UNSPSC Code:
41106300
NACRES:
NA.55

Quality Level

200

form

liquid

usage

sufficient for 100 reactions
 sufficient for 2000 reactions
 sufficient for 500 reactions

feature

dNTPs included
hotstart

technique(s)

qPCR: suitable

color

colorless

input

purified DNA

suitability

suitable for (quantitative PCR)

application(s)

agriculture

compatibility

Bio-Rad MyiQ ( )
 Bio-Rad iCycler iQ
 Bio-Rad iQ 5
for use with ABI 5700
for use with ABI 7000
for use with ABI 7300
for use with ABI 7500 Fast
for use with ABI 7500
for use with ABI 7700
for use with ABI 7900 Fast
for use with ABI 7900 HT
for use with ABI 7900
for use with ABI StepOne
for use with ABI StepOnePlus
for use with ABI ViiA 7
for use with Bio-Rad CFX384
for use with Bio-Rad CFX96
for use with Bio-Rad MJ Chromo4
for use with Bio-Rad MJ Opticon 2
for use with Bio-Rad MJ Opticon Cepheid SmartCycler
for use with Bio-Rad MJ Opticon
for use with Bio-Rad MiniOpticon
for use with Eppendorf® Mastercycler ep realplex2 s
for use with Eppendorf® Mastercycler ep realplex
for use with Illumina Eco qPCR
for use with Qiagen Corbett Rotor-Gene 3000
for use with Qiagen Corbett Rotor-Gene 6000
for use with Qiagen Corbett Rotor-Gene Q
for use with Roche LightCycler 480
for use with Strategene Mx3000P
for use with Strategene Mx3005P
for use with Strategene Mx4000

detection method

probe-based

shipped in

wet ice

storage temp.

−20°C

Categorías relacionadas

General description

Inclusion of JumpStart™ Taq antibody in the ReadyMix™ eliminates polymerase activity at ambient temperatures without causing the performance issues associated with chemically modified hot-start Taq. This allows for very rapid activation of the Taq and delivers unparalleled assay sensitivity, while allowing for benchtop reaction setup.

  • Kit is designed to perform probe-based qPCR assays on commonly available real-time instrument platforms.
  • ReadyMix requires the addition of reference dye (provided in kit) when being used on real-time instruments that require ROX passive reference dye for normalization of qPCR assays.
  • Kit is not compatible with qPCR instruments that utilize glass capillary tubes.
LuminoCt® qPCR ReadyMix combines the performance enhancements of our JumpStart™ Taq antibody for hot start PCR with the convenience of an easy-to-use reaction mixture.

Application

LuminoCt® qPCR ReadyMix™ has been used in 2-step quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay to quantify the reverse transcription product or cDNA for gene expression analysis. It has also been used in qPCR for quantifying the RNA extracted from rhinoviruses.

Features and Benefits

  • Ideal for rapid, high-throughput 2-step qPCR protocols
  • The master mix allows consistency and reproducibility from one reaction to the next
  • Reduced preparation time and the risk of contamination from multiple pipetting steps
  • Compatible with commonly available real-time instrument platforms for probe-based qPCR assays
  • Require small amplicons (<200 bp) for optimal results
  • Compatible with commercial primer sets, including TaqMan® assays
  • JumpStart™ Taq DNA polymerase allows room temperature set-up and the hot-start mechanism prevents primer-dimer and non-specific product formation
  • Assay results in as little as 25 minutes
  • Designed to deliver unsurpassed accuracy, precision, and sensitivity
  • Extensive design and validation of ReadyMix chemistry virtually eliminates the need to optimize assay parameters

Packaging

LuminoCt qPCR ReadyMix, Catalog Number L5794, contains optimized concentrations of Tris-HCl, pH 8.3, KCl, dNTPs (dATP, dCTP, dGTP, TTP), stabilizers, MgCl2 and JumpStart Taq DNA Polymerase. Provided as 100, 500 and 2000 reactions (25 μL mix in a 50 μL reaction volume).

100x ROX internal reference dye, Catalog Number R4526. Optional, for use with machines compatible with an internal reference dye, e.g., ABI and Stratagene.

Default reaction volume is 50 μL

100RXN is packaged as 1 X 2.5 mL
500RXN is packaged as 1 X 12.5 mL
2000RXN is packaged as 1 X 50 mL

Other Notes

  • Commercially available 5′ nuclease assays (e.g., TaqMan® Assays) are compatible with LuminoCt ReadyMix.
  • At a minimum, use software to design primers and probes for assays and ensure that qPCR amplicons are <200 bp.
  • Sigma recommends designing primers and probes with Beacon Designer software

To order primers and probes from Sigma, visit www.sigma.com/oligos

Legal Information

Eppendorf is a registered trademark of Eppendorf AG
JumpStart is a trademark of Sigma-Aldrich Co. LLC
LuminoCt is a registered trademark of Merck KGaA, Darmstadt, Germany
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
TaqMan is a registered trademark of Roche Molecular Systems, Inc.

Los componentes del kit también están disponibles por separado

Referencia del producto
Descripción
SDS
  • R4526100X ROX internal reference dyeSDS

Storage Class

10 - Combustible liquids

wgk_germany

WGK 1

Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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J Martínez et al.
Letters in applied microbiology, 50(6), 603-610 (2010-04-22)
To develop a real-time PCR-based strategy for the detection of Paenibacillus larvae vegetative cells and spores to improve the diagnosis and the screening of American foulbrood (AFB), the most harmful pathology of honeybee brood. A real-time PCR that allowed selective
Dieffenbach, C. and Dveksler, G., (eds)
PCR Primer: A Laboratory Manual (1995)
Marc J Champigny et al.
Frontiers in plant science, 4, 230-230 (2013-07-13)
DIR1 is a lipid transfer protein (LTP) postulated to complex with and/or chaperone a signal(s) to distant leaves during Systemic Acquired Resistance (SAR) in Arabidopsis. DIR1 was detected in phloem sap-enriched petiole exudates collected from wild-type leaves induced for SAR
Juan Vicente Canet et al.
The Plant cell, 24(10), 4220-4235 (2012-10-16)
Salicylic acid (SA) signaling acts in defense and plant development. The only gene demonstrated to be required for the response to SA is Arabidopsis thaliana non-expresser of pathogenesis-related gene 1 (NPR1), and npr1 mutants are insensitive to SA. By focusing
Philip Owens et al.
PloS one, 8(6), e67533-e67533 (2013-07-11)
Bone Morphogenetic Proteins (BMPs) are secreted cytokines that are part of the Transforming Growth Factor β (TGFβ) superfamily. BMPs have been shown to be highly expressed in human breast cancers, and loss of BMP signaling in mammary carcinomas has been
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Probe based QPCR utilizes a fluorescent–labeled target-specific probe resulting in increased specificity and sensitivity. Additionally, a variety of fluorescent dyes are available so that multiple primers can be used to simultaneously amplify many sequences.

PCR Master Mix

A PCR master mix is a batch of PCR or RT-PCR reagents that can be divided among many PCR reaction tubes. It usually includes DNA polymerase, dNTPs, MgCl2 and buffer. Make your own master mix or choose a commercial one.

Polymerase Chain Reaction

The polymerase chain reaction is one of the most widely used techniques in molecular biology. The PCR process consists of three main steps, Denaturation, Annealing & Extension

Protocolos

qPCR Gene Expression Using Probe Detection

The most common application for qPCR is the measurement of a gene transcript or copy number quantity relative to one or more reference genes using probe detection.

qPCR Using a Single Detection Probe Protocol

A protocol that can be used as a basic template for qPCR incorporating a detection probe that is specific to a single target. In these reactions, primers and probe are included at a final concentration of 200 nM and are run using LuminoCt® ReadyMix™.

Multiplex qPCR Protocol

A protocol that can be used as a basic template for qPCR incorporating up to four detection probes. In these reactions, primers and probe are included at a final concentration of 200 nM and are run using LuminoCt ReadyMix.

The 3'/5' Assay for Analysis of RNA Integrity Protocol

The 3’/5’ integrity assay is a potential first step in the identification of RNA degradation. The assay is particularly useful when a large number of samples are to be analyzed or when the degradation is less than that detected by capillary systems but still sufficient to effect qPCR analyses.

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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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