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Sigma-Aldrich

SYBR® Green Quantitative RT-qPCR Kit

One step SYBR® Green RT-qPCR with MMLV & hot-start Taq DNA Polymerase

NACRES:
NA.55

Quality Level

usage

sufficient for 250 reactions

feature

dNTPs included
hotstart

technique(s)

RT-qPCR: suitable

color

colorless

input

purified RNA

detection method

SYBR® Green

shipped in

wet ice

storage temp.

−20°C

Related Categories

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, and to detect mRNA expression levels.
The SYBR Green Quantitative RT-PCR kit provides a highly sensitive method for the quantitative analysis of gene expression. Sigma′s QRT-PCR ReadyMix combines the advantages of Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) and JumpStart Taq in a convenient ReadyMix.

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
  • JumpStartTaq 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.
JumpStart is a trademark of Sigma-Aldrich Co. LLC
SYBR is a registered trademark of Life Technologies

Kit Components Only

Product No.
Description

  • SYBR® Green Taq ReadyMix™ for Quantitative RT-PCR 2 x 25

Kit Components Also Available Separately

Product No.
Description
SDS

  • P219210X PCR Buffer, Optimized for routine PCR with MgCl2 included

  • M878725 mM MgCl2 1.5 mL/vial

  • R4526Reference Dye for Quantitative PCR, 100 ×, solution .3 mL/vial

Signal Word

Danger

Hazard Statements

Hazard Classifications

Aquatic Acute 1 - Aquatic Chronic 1 - Eye Dam. 1 - Skin Corr. 1C - Skin Sens. 1

Storage Class Code

8A - Combustible, corrosive hazardous materials

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

Gary P Richards et al.
Applied and environmental microbiology, 70(12), 7179-7184 (2004-12-03)
Genogroup I noroviruses from five genetic clusters and genogroup II noroviruses from eight genetic clusters were detected in stool extracts using degenerate primers and single-tube, real-time reverse transcription-PCR (RT-PCR) with SYBR Green detection. Two degenerate primer sets, designated MON 431-433
Pei Jin Lim et al.
PLoS neglected tropical diseases, 8(2), e2661-e2661 (2014-03-04)
Chikungunya virus (CHIKV) has resulted in several outbreaks in the past six decades. The clinical symptoms of Chikungunya infection include fever, skin rash, arthralgia, and an increasing incidence of encephalitis. The re-emergence of CHIKV with more severe pathogenesis highlights its
Validation of Quantitative PCR Assays
Lovatt, A., et al.
BioPharm., March, 22-23 (2002)
S A Bustin
Journal of molecular endocrinology, 29(1), 23-39 (2002-08-30)
The fluorescence-based real-time reverse transcription PCR (RT-PCR) is widely used for the quantification of steady-state mRNA levels and is a critical tool for basic research, molecular medicine and biotechnology. Assays are easy to perform, capable of high throughput, and can
T B Morrison et al.
BioTechniques, 24(6), 954-958 (1998-06-19)
Continuous fluorescence observation of amplifying DNA allows rapid and accurate quantification of initial transcript copy number. A simple and generic method for monitoring product synthesis with the double-stranded DNA dye, SYBR Green I provides initial template copy number estimation limited

Articles

Quantitative RT-PCR

RT-qPCR products combine the effective of Reverse Transcriptase with hot-start taq-directed antibody in convenient ReadyMixes for probe-based or SYBR® Green based applications.

Using probe-based quantitative PCR (qPCR) to measure gene-level expression

Quantitative PCR (qPCR) provides information about gene expression, gene amplification or loss, and small alterations. qPCR is often used to investigate tumor biology and to discover the genetic and epigenetic causes of cancer

Reverse Transcription

One approach to the analysis of gene expression is to measure the concentration of mRNA of a gene. There are several challenges to such analyses, such as the differences in half life between different transcripts, the temporal patterns of transcription and the lack of correlation between mRNA and protein.

PCR Master Mix

A PCR master mix is a batch of PCR or RT-PCR reagents that can be divided among many PCR reaction tubes. It usually includes DNA polymerase, dNTPs, MgCl2 and buffer. Make your own master mix or choose a commercial one.

Protocols

Primer Concentration Optimization Protocol

Primer Concentration Optimization Protocol is an approach to create a matrix of reactions. This is used to test a range of concentrations for each primer against different concentrations of the partner primer.

Reverse Transcription Protocol (One-step SYBR Green I Dye Detection)

Reverse transcription (RT) is the process of converting RNA to cDNA using a reverse transcriptase enzyme and dNTPs.

Coronavirus qPCR Design Case Study & Related Products to Support SARS-CoV-2 Research

Coronavirus qPCR Primer and probe sets for the detection of SARS-CoV-2 (Corona Virus), with additional real-time RT-PCR, RT-qPCR and supporting reagents available.

Related Content

RT-qPCR – Quantitative Reverse Transcription PCR

RT-qPCR, or quantitative reverse transcription PCR, combines the effects of reverse transcription and quantitative PCR or real-time PCR to amplify and detect specific targets. RT-qPCR has a variety of applications including quantifying gene expression levels, validating RNA interference (RNAi), and detecting pathogens such as viruses.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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