LuminoCt qPCR ReadyMix For fast probe-based quantitative PCR, 500 reactions

LuminoCt qPCR ReadyMix For fast probe-based quantitative PCR, 500 reactions

Synonym: LuminoCt qPCR ReadyMix, LuminoCt

•  NACRES NA.55

Properties

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. This is the ideal solution for performing high-throughput, quantitative PCR methods that rely on a fluorescent probe.

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.^[6][1][2]

LuminoCt qPCR ReadyMix is designed to deliver unsurpassed assay speeds without sacrificing accuracy, precision, or sensitivity. Extensive design and validation of the ReadyMix chemistry has virtually eliminated the need for optimization when LuminoCt is used in conjunction with properly designed primers and probes. 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.

Features and Benefits

• 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

• Assay results in as little as 25 minutes
• Unsurpassed accuracy, precision, and sensitivity
• Virtually eliminates the need to optimize assay parameters

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 μL

100RXN is packaged as 1 X 2.5 mL
500RXN is packaged as 1 X 12.5 mL
2000RXN is packaged as 1 X 50 mL

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 software

To order primers and probes from Sigma, visit www.sigma.com/oligos

Legal Information

Eppendorf is a registered trademark of Eppendorf AG

JumpStart is a trademark of Sigma-Aldrich Co. LLC

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

ReadyMix is a trademark of Sigma-Aldrich Co. LLC

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