Description
General description
The SYBR® Green Quantitative RT-PCR Kit combines Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT), JumpStart™ Taq DNA polymerase, and SYBR Green I fluorescent dye in a one-step RT-PCR kit designed for measurement of gene expression. This convenient 2X ready mix includes M-MLV RT, SYBR Green I dye, JumpStart™ Taq DNA polymerase, 99% pure deoxynucleotides, buffer, glass passivator, and stabilizers. The JumpStart™ 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
SYBR® Green Quantitative RT-qPCR Kit has been used in a 1-step reverse transcription polymerase chain reaction (RT-PCR) assay:to detect specific genetic clusters of genogroup I and II noroviruses
for chikungunya viral (CHIKV) RNA quantification
to detect mRNA expression levels
for amplification of total RNA extracted from human umbilical vein endothelial cells (HUVECs) and prostate cancer cells
The SYBR Green Quantitative RT-PCR Kit delivers high specificity and reproducibility. 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. Specificity of Sigma′s SYBR based QPCR detection is greatly enhanced by the incorporation of a hot-start mediated taq polymerase, JumpStart Taq. The kit has been optimized for use with both plate/tube real-time instruments and with the Roche LightCycler capillary instrument. A reference dye is provided in a separate vial to be used in ABI Detection Systems.
Features and Benefits
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
Reduced set-up time as compared to manual or wax Hot Start methods
JumpStart™Taq Polymerase reduces primer-dimer and non-specific product formation
SYBR® Green I dye is inexpensive, easy to use, and highly sensitive
Broad instrument compatibility
Includes a separate ROX reference dye vial for reaction normalization
Packaging
1 kit sufficient for 100 reactions at 50 μL each
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
JumpStart is a trademark of Sigma-Aldrich Co. LLC
SYBR is a registered trademark of Thermo Fisher Scientific or its subsidiaries
