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Sigma-Aldrich

Quantitative RT-PCR ReadyMix

One step RT-qPCR for probe-based methods, MMLV & hot-start Taq

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

probe-based

shipped in

wet ice

storage temp.

−20°C

Related Categories

General description

Quantitative RT-PCR ReadyMix combines Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) and JumpStart Taq DNA polymerase in a one-step RT-PCR kit designed for the measurement of gene expression. Tis convenient 2X ready mix includes M-MLV RT, 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. This kit has been formulated for fluorogenic hybridization probe-based detection methods.

Application

Quantitative RT-PCR ReadyMix has been used in the quantification of messenger RNA (mRNA) levels of specific expressed genes by quantitative reverse-transcription polymerase chain reaction (RT-qPCR).

Features and Benefits

  • The master mix allows consistency and reproducibility from one reaction to the next
  • Reduced 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
  • Broad instrument compatibility
  • Includes a separate ROX reference dye vial for reaction normalization

Packaging

1 kit sufficient for 250 reactions at 20 μL each or for 100 reactions at 50 μL each.

Legal Information

This product is for research use only.
JumpStart is a trademark of Sigma-Aldrich Co. LLC
ReadyMix is a trademark of Sigma-Aldrich Co. LLC

Kit Components Only

Product No.
Description

  • Probe Based qRT-PCR ReadyMix 2 X

  • Moloney Murine Leukemia Viral Reverse Transcriptase (M-MLV RT) 5000 U

Kit Components Also Available Separately

Product No.
Description
SDS

  • P219210X PCR Buffer, Optimized for routine PCR with MgCl2 included 1.5 mL/vial

  • M8787Magnesium chloride solution, PCR Reagent, 25 mM MgCI2 solution for PCR 25 mM

  • P219210X PCR Buffer, Optimized for routine PCR with MgCl2 included 100 X

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Skin Sens. 1

Storage Class Code

10 - Combustible liquids

Validation of Quantitative PCR Assays
Lovatt, A., et al.
BioPharm., 22-32 (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

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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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