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SYBR® Green Quantitative RT-qPCR Kit

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

Synonym(s):

one step rt qPCR, sybr green qPCR

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About This Item

NACRES:
NA.55
UNSPSC Code:
41106300

Key Documents

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usage

sufficient for 250 reactions

feature

dNTPs included, hotstart

technique(s)

RT-qPCR: suitable, qPCR: suitable

Quality Level

200

color

colorless

input

purified RNA

compatibility

ABI 5700, ABI 7000, ABI 7300, ABI 7500 Fast, ABI 7500, ABI 7700, ABI 7900 HT , ABI 7900 HT Fast, ABI 7900, ABI StepOne, ABI StepOnePlus, ABI ViiA 7, Bio-Rad CFX384, Bio-Rad CFX96, Bio-Rad MJ Chromo4, Bio-Rad MJ Opticon 2, Bio-Rad MJ Opticon, Bio-Rad MiniOpticon, Bio-Rad MyiQ, Bio-Rad iCycler iQ, Bio-Rad iQ5, Cepheid SmartCycler, Eppendorf® Mastercycler ep realplex2 s, Eppendorf® Mastercycler ep realplex, Illumina Eco qPCR, Qiagen Corbett Rotor-Gene 3000, Qiagen Corbett Rotor-Gene 6000, Qiagen Corbett Rotor-Gene Q, Roche LightCycler 480, Strategene Mx3000P, Strategene Mx3005P, Strategene Mx4000

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
  • 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

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 includedSDS
  • M878725 mM MgCl2 1.5 mL/vialSDS
  • R4526Reference Dye for Quantitative PCR, 100 ×, solution .3 mL/vialSDS

signalword

Danger

Storage Class

8A - Combustible corrosive hazardous materials

flash_point_f

Not applicable

flash_point_c

Not applicable

wgk

WGK 3

Hazard Classifications

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

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Certificates of Analysis (COA)

Lot/Batch Number
0000524118
0000524118
0000498397
0000498397
0000411010

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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
Carmela R Balistreri et al.
Scientific reports, 9(1), 11028-11028 (2019-08-01)
Bicuspid aortic valve (BAV) disease is recognized to be a syndrome with a complex and multifaceted pathophysiology. Its progression is modulated by diverse evolutionary conserved pathways, such as Notch-1 pathway. Emerging evidence is also highlighting the key role of TLR4
Phui San Ho et al.
Virology journal, 7, 13-13 (2010-01-23)
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus and one of the prevalent re-emerging arbovirus in tropical and subtropical regions of Asia, Africa, and Central and South America. It produces a spectrum of illness ranging from inapparent infection to moderate febrile
Caiyun Shan et al.
Molecular medicine reports, 17(4), 5300-5305 (2018-02-03)
Stroke is the most common cause of mortality worldwide. Post-stroke angiogenesis is of great significance to the treatment of strokes. The aim of the present study was to investigate the mechanism underlying the angiogenesis-promoting effect of microRNA‑126 (miR‑126)‑associated signaling pathways
Rong Ouyang et al.
Molecular medicine reports, 25(1) (2021-12-02)
Uric acid (UA) is the final oxidation product of purine metabolism. Hyperuricemia has been previously reported to contribute to vascular endothelial dysfunction and the development of cardiovascular diseases, metabolic syndrome and chronic kidney diseases. In addition, it has been reported
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Articles

PCR Master Mix

PCR master mix simplifies PCR/RT-PCR with components like DNA polymerase, dNTPs, MgCl2, and buffer, available commercially or DIY.

Probe QPCR for Gene-Level Expression

qPCR investigates gene expression, amplification, and alterations, crucial for tumor biology and understanding cancer genetics.

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.

Reverse Transcription

Challenges in gene expression analysis include mRNA stability, temporal transcription patterns, and mRNA-protein correlation, impacting accuracy.

Protocols

Reverse Transcription Protocol Using SYBR Green Dye Detection

Perform reverse transcription (RT) using a reverse transcriptase enzyme and dNTPs. Use total RNA or a gene-specific approach so that only the RNA of interest is converted to cDNA.

Primer Concentration Optimization

Primer Concentration Optimization Protocol creates reaction matrix for testing primer concentrations against various partners.

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