KiCqStart(R) One-Step Probe RT-qPCR ReadyMix(TM), ROX(TM)

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.