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KCQS07

Sigma-Aldrich

KiCqStart® One-Step Probe RT-qPCR ReadyMix

for Bio-Rad, Cepheid, Eppendorf, Illumina, Corbett, and Roche systems

Sinónimos:

KiCqStart® One Step Probe RT-qPCR Readymix

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About This Item

Código UNSPSC:
41106300
NACRES:
NA.55

Formulario

liquid

uso

sufficient for 500 reactions

Características

dNTPs included
hotstart

condiciones de almacenamiento

protect from light

técnicas

qPCR: suitable

color

light blue

entrada

purified RNA

compatibilidad

Bio-Rad CFX384
Bio-Rad CFX96
Bio-Rad MJ Chromo4
Bio-Rad MJ Opticon 2
Bio-Rad MJ Opticon
Bio-Rad MiniOpticon
Eppendorf® Mastercycler ep realplex2 s
Eppendorf® Mastercycler ep realplex
Illumina Eco qPCR
Qiagen Corbett Rotor-Gene 3000
Qiagen Corbett Rotor-Gene 6000
Qiagen Corbett Rotor-Gene Q
Roche LightCycler 480

método de detección

probe-based

Condiciones de envío

dry ice

temp. de almacenamiento

−20°C

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Descripción general

KiCqStart® One-Step Probe RT-qPCR ReadyMix is a ready-to-use, highly sensitive master mix for reverse transcription quantitative PCR (RT-qPCR) of RNA templates using hybridization probe detection chemistries such as TaqMan® 5′-hydrolysis probes on real-time PCR systems (refer to instrument compatibility section to select the appropriate reagent for your machine). First-strand cDNA synthesis and PCR amplification are carried out in the same tube without opening between procedures. It is ideal for highly sensitive quantification of RNA viruses or low abundance RNA targets as well as high throughput gene-expression studies. The system has been optimized to deliver maximum RT-PCR efficiency, sensitivity, and specificity in reduced reaction volumes and fast cycle times.

KiCqStart® One-Step Probe RT-qPCR ReadyMix contains all required components for RT-qPCR except RNA template and probe. It is compatible with all dual-labeled probe chemistries. The reverse transcriptase is an engineered M-MLV with reduced RNase H activity and improved activity and stability at higher temperatures. The use of higher temperatures (50 to 55°C) for the first-strand step of one-step RT-qPCR provides higher specificity for primer annealing and disruption of RNA secondary structure that can interfere with cDNA synthesis.

KiCqStart® One-Step Probe RT-qPCR ReadyMix is highly resistant to PCR inhibitors. A key component of the ReadyMix is an ultra-pure, highly processive, thermostable DNA polymerase that is combined with high avidity monoclonal antibodies. This provides an extremely stringent automatic hot-start that minimizes the potential for primer-dimer and other non-specific PCR artifacts. The light blue color of the inert dye simplifies reaction assembly in white, or clear, plates and helps to minimize pipetting or mixing errors. The dye does not interfere with qPCR performance or affect the stability of the product.

Aplicación

KiCqStart® One-Step Probe RT-qPCR ReadyMix has been used:
  • in quantitative real-time polymerase chain reaction (qRT-PCR) for gene expression studies
  • in cell type-specific transcript analysis for determining the cycle threshold (Ct) value
  • for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA detection from nasopharyngealand oropharyngeal swabs
  • for amplification of reverse-transcribed cDNA

Características y beneficios

  • Assay results in as little as 33 minutes
  • Highly efficient and sensitive real-time PCR results
  • Little/no optimization required

Componentes

2X reaction buffer containing dATP, dCTP, dGTP, dTTP, magnesium chloride, reverse transcriptase, RNase inhibitor protein, hot-start DNA polymerase, inert blue qPCR dye, and stabilizers

packaging:
500 reactions* = 5 X 1 mL tubes
*number of reactions based on a 20uL volume

Otras notas

Different real-time PCR systems employ different strategies for normalization of fluorescent signals and correction of well-to-well optical variations. It is critical to match the appropriate qPCR reagent to your specific instrument.KiCqStart® One-Step Probe RT-qPCR ReadyMix does not contain an internal reference dye. Compatible instruments include:
  • Bio-Rad® CFX384
  • Bio-Rad CFX96
  • Bio-Rad MiniOpticon
  • Bio-Rad/MJ Chromo4
  • Bio-Rad/MJ Opticon 2
  • Bio-Rad/MJ Opticon
  • Cepheid SmartCycler®
  • Eppendorf Mastercycler® ep realplex
  • Eppendorf Mastercycler ep realplex2 s
  • Illumina Eco qPCR
  • Qiagen/Corbett Rotor-Gene® 3000
  • Qiagen/Corbett Rotor-Gene 6000• Qiagen/Corbett Rotor-Gene Q
  • Roche LightCycler® 480

Información legal

Bio-Rad is a registered trademark of Bio-Rad Laboratories, Inc.
CFX384 is a trademark of Bio-Rad Laboratories, Inc.
CFX96 is a trademark of Bio-Rad Laboratories, Inc.
Chromo4 is a trademark of Bio-Rad Laboratories, Inc.
Eppendorf is a registered trademark of Eppendorf AG
KiCqStart is a registered trademark of QIAGEN Beverly Inc.
LightCycler is a registered trademark of Roche
Mastercycler is a registered trademark of Eppendorf AG
MiniOpticon is a trademark of Bio-Rad Laboratories, Inc.
Opticon is a trademark of Bio-Rad Laboratories, Inc.
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
Rotor-Gene is a registered trademark of Qiagen GmbH
SmartCycler is a registered trademark of Cepheid, Inc.
TaqMan is a registered trademark of Roche Molecular Systems, Inc.

Código de clase de almacenamiento

12 - Non Combustible Liquids

Clase de riesgo para el agua (WGK)

WGK 3

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable


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Visite la Librería de documentos

Myonuclear transcription is responsive to mechanical load and DNA content but uncoupled from cell size during hypertrophy
Kirby T J, et al.
Molecular Biology of the Cell, 27(5), 788-798 (2016)
Integrative mRNA-microRNA analyses reveal novel interactions related to insulin sensitivity in human adipose tissue
Kirby T J, et al.
Physiological Genomics, 48(2), 145-153 (2015)
Insulin-resistant subjects have normal angiogenic response to aerobic exercise training in skeletal muscle, but not in adipose tissue
Walton R G, et al.
Physiological Reports, 3(6), 145-153 (2015)
R Grace Walton et al.
Physiological reports, 3(6), doi:10-doi:10 (2015-06-04)
Reduced vessel density in adipose tissue and skeletal muscle is associated with obesity and may result in decreased perfusion, decreased oxygen consumption, and insulin resistance. In the presence of VEGFA, Angiopoietin-2 (Angpt2) and Angiopoietin-1 (Angpt1) are central determinants of angiogenesis
Tyler J Kirby et al.
Physiological genomics, 48(2), 145-153 (2015-12-17)
Adipose tissue has profound effects on whole-body insulin sensitivity. However, the underlying biological processes are quite complex and likely multifactorial. For instance, the adipose transcriptome is posttranscriptionally modulated by microRNAs, but the relationship between microRNAs and insulin sensitivity in humans

Artículos

Sigma’s RT-qPCR products combine the effective of Reverse Transcriptase with hot-start taq-directed antibody in convenient ReadyMixes for probe-based or SYBR® Green based applications.

PCR assay guide navigates you through primer validation and other assay optimization factors to ensure high sensitivity and specificity for optimum DNA/ RNA quantification.

One approach to the analysis of gene expression is to measure the concentration of mRNA of a gene. There are several challenges to such analyses, such as the differences in half life between different transcripts, the temporal patterns of transcription and the lack of correlation between mRNA and protein.

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.

Ver todo

Protocolos

KiCqStart One-Step Probe RT-qPCR ReadyMix is a ready-to-use, highly sensitive master mix for reverse transcription quantitative PCR (RT-qPCR) of RNA templates using hybridization probe detection chemistries such as TaqMan 5'-hydrolysis probes on real-time PCR systems.

Coronavirus qPCR Primer and probe sets for the detection of SARS-CoV-2 (Corona Virus), with additional real-time RT-PCR, RT-qPCR and supporting reagents available.

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