KiCqStart(R) One-Step Probe RT-qPCR ReadyMix(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

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

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, inert blue qPCR dye, and stabilizers

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

Other Notes

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

Legal Information

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.