Description
General description
SYBR® Green JumpStart™ Taq ReadyMix™ combines the performance enhancements of JumpStart Taq antibody for hot start PCR with SYBR Green I and the convenience of an easy-to-use ReadyMix solution. This ready-to-use mixture of SYBR Green I, JumpStart Taq DNA polymerase, 99% pure deoxynucleotides and reaction buffer is provided in a 2× concentrate for ease of use. Simply add 25 μL of the 2× mix to DNA template, primers and water. The JumpStart Taq antibody inactivates DNA polymerase at room temperature. When the temperature is raised above 70 °C in the first denaturation step of the cycling process, the complex dissociates and the polymerase becomes fully active. JumpStart Taq DNA polymerase prevents non-specific amplification resulting in more accurate CT values.
Application
SYBR® Green JumpStart™ Taq ReadyMix™ has been used:
to prepare the real-time PCR mixture
in the quantitative analysis of gene expression by real time, quantitative polymerase chain reaction (qPCR)
to determine the mRNA expression of nuclear factor erythroid 2 (NFE2)-related factor 2 (Nrf2) gene by qPCR
SYBR® Green JumpStart™ Taq ReadyMix™ has been used:
During RNA extraction, reverse transcription and Quantitative PCR
During analysis of leptin and leptin receptor expression. (PCR and melting curve analyses were performed)
For tissue RNA Isolation. (PCR and melting curve analyses were performed)
in real-time PCR (RT-PCR) of reverse-transcribed cDNA template
For qPCR amplifications using SYBR
For multiplex PCR
Biochem/physiol Actions
SYBR Green JumpStart Taq ReadyMix is recommended for single product real-time amplification experiments and can also be used for PCR optimization prior to manufacture of fluorescent-labeled probes. Fluorescent labeled probes are not recommended for use with SYBR Green I dye.
SYBR Green I, a commonly used fluorescent DNA binding dye, binds all double-stranded DNA and detection is monitored by measuring the increase in fluorescence throughout the cycle. SYBR Green I has an excitation and emission maxima of 494 nm and 521 nm, respectively. The instrument settings for ROX reference dye are satisfactory for the measurement of the Reference Dye for Quantitative PCR. Specificity of Sigma′s SYBR based QPCR detection is greatly enhanced by the incorporation of a hot-start mediated taq polymerase, JumpStart Taq.
Features and Benefits
Delivers the benefits of antibody-inactivated hot-start PCR with SYBR Green detection in a ReadyMix ideal for high throughput applications; only primers and template are required
JumpStart™ Taq DNA polymerase prevents amplification of non-specific products, resulting in increased efficiency and higher target yield
SYBR® Green JumpStart™ Taq ReadyMix for SYBR based qPCR is formulated with MgCl₂ or packaged with a separate vial for ease of optimization. ReadyMixes are compatible with tube- and plate-based instruments
This master mix allows consistency from one reaction to the next
Designed to minimize contamination
Reduced primer dimers
Reduced set-up time as compared to manual or wax HotStart® methods
Allows for room temperature set-up and contains a fluorescent dye, which is ideal for qPCR applications
Packaging
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
500RXN is packaged as 1 X 12.5 mL
Other Notes
Magnesium chloride is provided separately for optimization.
Legal Information
Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,994,056 and 6,171,785.. 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. No right under any other patent claim (such as apparatus or system claims in US Patent No. 6,814,934) 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 by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
Eppendorf is a registered trademark of Eppendorf SE
HOTSTART is a registered trademark of Molecular BioProducts, Inc.
JumpStart is a trademark of Sigma-Aldrich Co. LLC
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
SYBR is a registered trademark of Thermo Fisher Scientific or its subsidiaries
