S5193

SYBR^® Green JumpStart^™ Taq ReadyMix^™

for quantitative PCR, without MgCl₂ in buffer

• Synonym(s): Hot start PCR master mix, sybr green PCR master mix, sybr green qPCR
• MDL number: MFCD03458666
• UNSPSC Code: 41106300
• NACRES: NA.55

Properties

• Quality Level: 200
• form: liquid
• usage: sufficient for 100 reactions; sufficient for 20 reactions; sufficient for 400 reactions
• feature: dNTPs included; hotstart
• concentration: 1.25 units/reaction (50 μL reaction volume)
• technique(s): qPCR: suitable
• color: colorless
• input: purified DNA
• suitability: suitable for PCR
• application(s): agriculture
• compatibility: Bio-Rad MyiQ; Bio-Rad iCycler iQ; Bio-Rad iQ 5; for use with ABI 5700; for use with ABI 7000; for use with ABI 7300; for use with ABI 7500 Fast; for use with ABI 7500; for use with ABI 7700; for use with ABI 7900 Fast; for use with ABI 7900 HT; for use with ABI 7900; for use with ABI StepOne; for use with ABI StepOnePlus; for use with ABI ViiA 7; for use with Bio-Rad CFX384; for use with Bio-Rad CFX96; for use with Bio-Rad MJ Chromo4; for use with Bio-Rad MJ Opticon 2; for use with Bio-Rad MJ Opticon Cepheid SmartCycler; for use with Bio-Rad MJ Opticon; for use with Bio-Rad MiniOpticon; for use with Eppendorf^® Mastercycler ep realplex2 s; for use with Eppendorf^® Mastercycler ep realplex; for use with Illumina Eco qPCR; for use with Qiagen Corbett Rotor-Gene 3000; for use with Qiagen Corbett Rotor-Gene 6000; for use with Qiagen Corbett Rotor-Gene Q; for use with Roche LightCycler 480; for use with Strategene Mx3000P; for use with Strategene Mx3005P; for use with Strategene Mx4000
• detection method: SYBR^® Green
• shipped in: wet ice
• storage temp.: −20°C

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 C_T values.

Application

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

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

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.

Principle

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.

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 AG

HOTSTART is a registered trademark of Molecular BioProducts, Inc.

Julabo is a registered trademark of Julabo USA, 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 Life Technologies

Safety Information

• Pictograms: GHS07,GHS09
• Signal Word: Warning
• Hazard Statements: H315,H317,H319,H410
• Precautionary Statements: P261 - P264 - P273 - P280 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Aquatic Acute 1 - Aquatic Chronic 1 - Eye Irrit. 2 - Skin Irrit. 2 - Skin Sens. 1
• Storage Class Code: 10 - Combustible liquids
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable