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KCQS09

Sigma-Aldrich

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

with ROX for ABI instruments

Synonym(s):

RT qPCR, one step RT qPCR, probe PCR

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About This Item

UNSPSC Code:
41106300
NACRES:
NA.55

form

liquid

usage

sufficient for 500 reactions

feature

dNTPs included
hotstart

storage condition

protect from light

technique(s)

RT-qPCR: suitable

color

light blue

input

purified RNA

compatibility

ABI 5700
ABI 7000
ABI 7300
ABI 7700
ABI 7900 HT
ABI 7900 HT Fast
ABI 7900
ABI StepOne
ABI StepOnePlus

detection method

probe-based

shipped in

dry ice

storage temp.

−20°C

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® 5700
  • Applied Biosystems 7000
  • Applied Biosystems 7300
  • Applied Biosystems 7700
  • Applied Biosystems 7900
  • Applied Biosystems 7900 HT Fast
  • Applied Biosystems 7900 HT
  • Applied Biosystems StepOnePlus
  • Applied Biosystems StepOne

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

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.
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
StepOne is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
StepOnePlus is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
TaqMan is a registered trademark of Roche Molecular Systems, Inc.

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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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Articles

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.

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

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.

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.

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.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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