Application

LuminoCt^® qPCR ReadyMix^™ has been used in 2-step quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay to quantify the reverse transcription product or cDNA for gene expression analysis. It has also been used in qPCR for quantifying the RNA extracted from rhinoviruses.

Features and Benefits

Assay results in as little as 25 minutes Designed to deliver unsurpassed accuracy, precision, and sensitivity Extensive design and validation of ReadyMix chemistry virtually eliminates the need to optimize assay parameters

Ideal for rapid, high-throughput 2-step qPCR protocols The master mix allows consistency and reproducibility from one reaction to the next Reduced preparation time and the risk of contamination from multiple pipetting steps Compatible with commonly available real-time instrument platforms for probe-based qPCR assays Require small amplicons (﹤200 bp) for optimal results Compatible with commercial primer sets, including TaqMan^® assays JumpStart^™ Taq DNA polymerase allows room temperature set-up and the hot-start mechanism prevents primer-dimer and non-specific product formation

General description

LuminoCt^® qPCR ReadyMix combines the performance enhancements of our JumpStart^™ Taq antibody for hot start PCR with the convenience of an easy-to-use reaction mixture.

Inclusion of JumpStart^™ Taq antibody in the ReadyMix^™ eliminates polymerase activity at ambient temperatures without causing the performance issues associated with chemically modified hot-start Taq. This allows for very rapid activation of the Taq and delivers unparalleled assay sensitivity, while allowing for benchtop reaction setup. Kit is designed to perform probe-based qPCR assays on commonly available real-time instrument platforms. ReadyMix requires the addition of reference dye (provided in kit) when being used on real-time instruments that require ROX passive reference dye for normalization of qPCR assays. Kit is not compatible with qPCR instruments that utilize glass capillary tubes.

Legal Information

TaqMan is a registered trademark of Roche Molecular Systems, Inc.

JumpStart is a trademark of Sigma-Aldrich Co. LLC

ReadyMix is a trademark of Sigma-Aldrich Co. LLC

Eppendorf is a registered trademark of Eppendorf AG

LuminoCt is a registered trademark of Sigma-Aldrich Co. LLC

Other Notes

Commercially available 5′ nuclease assays (e.g., TaqMan^® Assays) are compatible with LuminoCt ReadyMix. At a minimum, use software to design primers and probes for assays and ensure that qPCR amplicons are ﹤200 bp. Sigma recommends designing primers and probes with Beacon Designer softwareTo order primers and probes from Sigma, visit www.sigma.com/oligos

Packaging

LuminoCt qPCR ReadyMix, Catalog Number L5794, contains optimized concentrations of Tris-HCl, pH 8.3, KCl, dNTPs (dATP, dCTP, dGTP, TTP), stabilizers, MgCl₂ and JumpStart Taq DNA Polymerase. Provided as 100, 500 and 2000 reactions (25 µL mix in a 50 µL reaction volume).100x ROX internal reference dye, Catalog Number R4526. Optional, for use with machines compatible with an internal reference dye, e.g., ABI and Stratagene.Default reaction volume is 50 µL100RXN is packaged as 1 X 2.5 mL500RXN is packaged as 1 X 12.5 mL2000RXN is packaged as 1 X 50 mL