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KCQS07

Sigma-Aldrich

KiCqStart® One-Step Probe RT-qPCR ReadyMix

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

Synonyme(s) :

KiCqStart® One Step Probe RT-qPCR Readymix

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About This Item

Code UNSPSC :
41106300
Nomenclature NACRES :
NA.55

Forme

liquid

Utilisation

sufficient for 500 reactions

Caractéristiques

dNTPs included
hotstart

Conditions de stockage

protect from light

Technique(s)

qPCR: suitable

Couleur

light blue

Entrée

purified RNA

Compatibilité

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

Méthode de détection

probe-based

Conditions d'expédition

dry ice

Température de stockage

−20°C

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Description générale

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.

Application

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

Caractéristiques et avantages

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

Composants

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

Autres remarques

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

Informations légales

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.

Code de la classe de stockage

12 - Non Combustible Liquids

Classe de danger pour l'eau (WGK)

WGK 3

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable


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Certificats d'analyse (COA)

Lot/Batch Number

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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.

Consulter la Bibliothèque de documents

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

Articles

Viral RNA Extraction Buffer (VRE100) and related reagents for qRT-PCR and additional viral RNA analyses.

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.

Afficher tout

Protocoles

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.

Contenu apparenté

GenElute™-E Single Spin DNA and RNA purification technology enables rapid DNA and RNA extraction and purification from mammalian cells, blood, tissue, and plant tissue samples.

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.

Notre équipe de scientifiques dispose d'une expérience dans tous les secteurs de la recherche, notamment en sciences de la vie, science des matériaux, synthèse chimique, chromatographie, analyse et dans de nombreux autres domaines..

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