MilliporeSigma
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D7440

Sigma-Aldrich

JumpStart Taq ReadyMix for Quantitative PCR

For probe-based real-time PCR

Synonym(s):
hot start PCR, hot start master mix, hot start DNA polymerase, qPCR, real time quantitative PCR
NACRES:
NA.55

form

liquid

usage

sufficient for 100 reactions
sufficient for 20 reactions
sufficient for 400 reactions

feature

Multiplex PCR
dNTPs included
hotstart

concentration

1.5 units/reaction (50 μL reaction volume)

technique(s)

qPCR: suitable

color

colorless

input

purified DNA

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

probe-based

shipped in

wet ice

storage temp.

−20°C

General description

JumpStart Taq ReadyMix is a ready-to-use 2X master mix that contains JumpStart Taq DNA polymerase, 99% pure dNTPs, reaction buffer and JumpStart Taq antibody. It combines the performance enhancements of our JumpStart Taq Antibody for hot start PCR with the convenience of an easy-to-use reaction mixture. Since it has no added dyes, this is the ideal solution for performing high-throughput, quantitative PCR methods that rely on a fluorescent probe. The 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

JumpStart Taq ReadyMix for Quantitative PCR has been used:
  • for quantitative polymerase chain reaction (qPCR) amplification of HIV-1 RNA that is reverse transcribed and DNA using probes
  • in a 2-step RT-qPCR assay for the quantification of reverse transcribed RNA in a study to monitor the activity of licorice in Treg cell differentiation and function
  • as a component of the reaction mixture for the detection and amplification of Salmonella sp. and internal amplification control (IAC) pUC 19 plasmid DNA
  • as a component of the reaction mixture for the detection of Clostridium difficile by quantitative polymerase chain reaction (qPCR)
  • for quantitative real-time -PCR of reverse-transcribed ß-arrestin-1 (ARRB1) and β-arrestin-2 (ARRB2) mRNA

Features and Benefits

  • Ultimate Convenience: ReadyMixes contain all components necessary for QPCR, simply add fluorescent detection chemistry, primers, and template
  • Greater Specificity & Increased Target Yield: JumpStart Taq prevents non-specific amplification and increased target yield
  • Maximum Flexibility: Suitable for use with a variety of detection chemistries including molecular probes and double-stranded binding dyes such as SYBR® Green I
  • Designed for use with either plate/tube real-time thermal cyclers or capillary instruments

Packaging

Sigma′s Reference Dye for Quantitative PCR is included separately with this ReadyMix for normalization of the reaction data. The dye has a maximum excitation of 586 nm, and a maximum emission of 605 nm. The instrument settings for ROX reference dye are satisfactory for the measurement of the Reference Dye for Quantitative PCR. A tube of 25 mM MgCl2 is also provided for easy optimization of the qPCR reaction.

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

Other Notes

JumpStart Taq ReadyMix for Quantitative PCR combines the advantages of a hot start enzyme with a ready-to-use mix for high throughput, quantitative PCR (qPCR). It is formulated without a detection chemistry making it suitable for use with a variety of formats including dual-labeled probes, molecular beacons, or double stranded binding dyes such as SYBR Green I.

Principle

The ReadyMix contains JumpStart Taq DNA Polymerase, 99% pure deoxynucleotides, and buffer in an optimized 2× concentrate. To prepare a reaction, 25 μL of the ReadyMix is added to primers, template, detection chemistry, and water for a total reaction volume of 50 μL. Set up can be performed at room temperature since the JumpStart Taq antibody renders the Taq DNA polymerase inactive. During the first denaturation cycle, the antibody dissociates from the enzyme and full activity is restored. No special preparations or protocol changes are required.

Unit Definition

One unit incorporates 10 nmol of total dNTPs into acid-precipitable DNA in 30 min. at 74 °C.

Legal Information

A license to perform the patented 5′ Nuclease Process for research is obtained by the purchase of (i) both Authorized 5′ Nuclease Core Kit and Licensed Probe, (ii) a Licensed 5′ Nuclease Kit, or (iii) license rights from Applied Biosystems.

This product is an Authorized 5′ Nuclease Core Kit. Use of this product is covered by one or more of the following claims outside the US corresponding to US Patent No. 5,210,015 and 5,487,972. 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. Separate purchase of a Licensed Probe would convey rights under US Patents and corresponding patent claims outside the US: 5,538,848, 5,723,591, 5,876,930, 6,030,787, 6,258,569, 5,804,375 (claims 1-12 only), and and claims outside the United States corresponding to US Patent No. 6,214,979. No right under any other patent claim 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 from the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
Eppendorf is a registered trademark of Eppendorf AG
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

Kit Components Also Available Separately

Product No.
Description
SDS

  • P2893JumpStart Taq ReadyMix, Complete optimized reagent for hot-start PCR at 2X concentration 2 X

  • M878725 mM MgCl2 1.5 μL

  • R4526Reference Dye for Quantitative PCR, 100 ×, solution 100 X

Pictograms

Exclamation markEnvironment

Signal Word

Warning

Hazard Statements

Hazard Classifications

Aquatic Acute 1 - Aquatic Chronic 1 - Eye Irrit. 2 - Skin Irrit. 2 - Skin Sens. 1

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

J G Schmidt et al.
Fish & shellfish immunology, 48, 273-284 (2015-12-26)
We followed the progression of healing of deep excisional biopsy punch wounds over the course of 365 days in rainbow trout (Oncorhynchus mykiss) by monitoring visual wound healing and gene expression in the healing muscle at regular intervals (1, 3
Andrew J Levine et al.
Journal of neurovirology, 22(3), 366-375 (2015-12-23)
HIV infection leads to age-related conditions in relatively young persons. HIV-associated neurocognitive disorders (HAND) are considered among the most prevalent of these conditions. To study the mechanisms underlying this disorder, researchers need an accurate method for measuring biological aging. Here
Benjamin B Gelman et al.
Journal of acquired immune deficiency syndromes (1999), 62(5), 487-495 (2012-12-18)
Replicating HIV-1 in the brain is present in HIV encephalitis (HIVE) and microglial nodule encephalitis (MGNE) and is putatively linked with HIV-associated neurocognitive disorders (HAND). A cliniconeurovirological correlation was conducted to elucidate the relationship between brain viral load and clinical
Cawas B Engineer et al.
Plant physiology, 143(1), 236-250 (2006-11-07)
Nitrogen is an essential macronutrient for plant growth and survival. Here, the temporal and spatial sensing of nitrogen starvation is analyzed in Arabidopsis (Arabidopsis thaliana). The promoter for the high-affinity ammonium transporter, AtAmt1.1, is shown to be a valid indicator
Lithium, an anti-psychotic drug, greatly enhances the generation of induced pluripotent stem cells.
Wang, Q., et al.
Cell Research (2011)

Articles

Multiplex qPCR with JumpStart™ Taq ReadyMix™ for Quantitative PCR

Method outlines use of a hot start Taq for multiplex qPCR and provides guidance on how to optimize dNTPs, primer, probes and MgCL2 concentrations. By optimizing these parameters, the user can improve assay sensitivity and linear range of detection.

Probe-based qPCR

Probe based QPCR utilizes a fluorescent–labeled target-specific probe resulting in increased specificity and sensitivity. Additionally, a variety of fluorescent dyes are available so that multiple primers can be used to simultaneously amplify many sequences.

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

Polymerase Chain Reaction

The polymerase chain reaction is one of the most widely used techniques in molecular biology. The PCR process consists of three main steps, Denaturation, Annealing & Extension

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