KCQS08

Sigma-Aldrich

KiCqStart® One-Step Probe RT-qPCR ReadyMix, Low ROX

with Low ROX for ABI and Stratagene instruments

Synonym(s):
KiCqStart One Step Probe RT-qPCR Readymix
NACRES:
NA.55
Pricing and availability is not currently available.

form

liquid

feature

hotstart

storage condition

protect from light

color

light blue

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description

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

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:
  • Applied Biosystems® 7500
  • Applied Biosystems 7500 Fast
  • Applied Biosystems ViiA 7
  • Applied Biosystems QuantStudio
  • Agilent® AriaMx
  • Douglas Scientific IntelliQube
  • QIAGEN® Rotor-Gene® Q
  • Stratagene Mx3000P®
  • Stratagene Mx3005P®
  • Stratagene Mx4000

Features and Benefits

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

Components

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

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

Legal Information

Agilent is a registered trademark of Agilent Technologies, Inc.
Applied Biosystems is a registered trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
KiCqStart is a registered trademark of Qiagen Beverly Inc.
Mx3000P is a registered trademark of Agilent Technologies, Inc.
Mx3005P is a registered trademark of Agilent Technologies, Inc.
Mx4000 is a trademark of Agilent Technologies, Inc.
Qiagen is a registered trademark of Qiagen Group
ROX is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
Rotor-Gene is a registered trademark of Qiagen GmbH
TaqMan is a registered trademark of Roche Molecular Systems, Inc.

RIDADR

NONH for all modes of transport

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

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.
Molecular biology of the cell, 27(5), 788-798 (2016-01-15)
Myofibers increase size and DNA content in response to a hypertrophic stimulus, thus providing a physiological model with which to study how these factors affect global transcription. Using 5-ethynyl uridine (EU) to metabolically label nascent RNA, we measured a sevenfold...
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
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.
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PCR assay guide navigates you through primer validation and other assay optimization factors to ensure high sensitivity and specificity for optimum DNA/ RNA quantification.
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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.
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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
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Protocols
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.
Read More
Related Content
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.
Read More

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