JumpStart(TM) Taq ReadyMix(TM) for Quantitative PCR (C15-1355-873)

General description

JumpStart^™ Taq ReadyMix^™ is a ready-to-use 2X master mix that contains JumpStart^™ Taq DNA polymerase, 99% pure dNTPs, reaction buffer and JumpStart^™ Taq antibody. It combines the performance enhancements of our JumpStart^™ Taq Antibody for hot start PCR with the convenience of an easy-to-use reaction mixture. Since it has no added dyes, this is the ideal solution for performing high-throughput, quantitative PCR methods that rely on a fluorescent probe. The Taq DNA polymerase is an antibody-inactivated hot-start enzyme. Once the reaction temperature reaches 70°C, the DNA polymerase-antibody complex dissociates and Taq DNA polymerase activity is restored. This antibody-enzyme complex allows for easy and convenient set-up with less contamination risk than manual hot-start techniques.

Application

JumpStart^™ Taq ReadyMix^™ for Quantitative PCR has been used:

• for quantitative polymerase chain reaction (qPCR) amplification of HIV-1 RNA that is reverse transcribed and DNA using probes
• in a 2-step RT-qPCR assay for the quantification of reverse transcribed RNA in a study to monitor the activity of licorice in Treg cell differentiation and function
• as a component of the reaction mixture for the detection and amplification of Salmonella sp. and internal amplification control (IAC) pUC 19 plasmid DNA
• as a component of the reaction mixture for the detection of Clostridium difficile by quantitative polymerase chain reaction (qPCR)
• for quantitative real-time -PCR of reverse-transcribed ß-arrestin-1 (ARRB1) and β-arrestin-2 (ARRB2) mRNA

Features and Benefits

• Ultimate Convenience: ReadyMixes contain all components necessary for QPCR, simply add fluorescent detection chemistry, primers, and template
• Greater Specificity & Increased Target Yield: JumpStart^™ Taq prevents non-specific amplification and increased target yield
• Maximum Flexibility: Suitable for use with a variety of detection chemistries including molecular probes and double-stranded binding dyes such as SYBR^® Green I
• Designed for use with either plate/tube real-time thermal cyclers or capillary instruments

Packaging

Sigma′s Reference Dye for Quantitative PCR is included separately with this ReadyMix for normalization of the reaction data. The dye has a maximum excitation of 586 nm, and a maximum emission of 605 nm. The instrument settings for ROX reference dye are satisfactory for the measurement of the Reference Dye for Quantitative PCR. A tube of 25 mM MgCl₂ is also provided for easy optimization of the qPCR reaction.

Default reaction volume is 50 μL

20RXN is packaged as 1 X 500 μL
100RXN is packaged as 1 X 2.5 mL
400RXN is packaged as 1 X 10 mL

Other Notes

JumpStart Taq ReadyMix for Quantitative PCR combines the advantages of a hot start enzyme with a ready-to-use mix for high throughput, quantitative PCR (qPCR). It is formulated without a detection chemistry making it suitable for use with a variety of formats including dual-labeled probes, molecular beacons, or double stranded binding dyes such as SYBR Green I.

Principle

The ReadyMix contains JumpStart Taq DNA Polymerase, 99% pure deoxynucleotides, and buffer in an optimized 2× concentrate. To prepare a reaction, 25 μL of the ReadyMix is added to primers, template, detection chemistry, and water for a total reaction volume of 50 μL. Set up can be performed at room temperature since the JumpStart Taq antibody renders the Taq DNA polymerase inactive. During the first denaturation cycle, the antibody dissociates from the enzyme and full activity is restored. No special preparations or protocol changes are required.

Unit Definition

One unit incorporates 10 nmol of total dNTPs into acid-precipitable DNA in 30 min. at 74 °C.

Legal Information

A license to perform the patented 5′ Nuclease Process for research is obtained by the purchase of (i) both Authorized 5′ Nuclease Core Kit and Licensed Probe, (ii) a Licensed 5′ Nuclease Kit, or (iii) license rights from Applied Biosystems.

This product is an Authorized 5′ Nuclease Core Kit. Use of this product is covered by one or more of the following claims outside the US corresponding to US Patent No. 5,210,015 and 5,487,972. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. Separate purchase of a Licensed Probe would convey rights under US Patents and corresponding patent claims outside the US: 5,538,848, 5,723,591, 5,876,930, 6,030,787, 6,258,569, 5,804,375 (claims 1-12 only), and and claims outside the United States corresponding to US Patent No. 6,214,979. No right under any other patent claim and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained from the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Eppendorf is a registered trademark of Eppendorf AG

JumpStart is a trademark of Sigma-Aldrich Co. LLC

ReadyMix is a trademark of Sigma-Aldrich Co. LLC

SYBR is a registered trademark of Life Technologies