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Sigma-Aldrich

Quantitative RT-PCR ReadyMix™

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

Synonym(s):

1-step RT-qPCR mix, Quantitative real-time PCR master mix

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About This Item

UNSPSC Code:
41106300
NACRES:
NA.55

usage

sufficient for 250 reactions

feature

dNTPs included
hotstart

technique(s)

RT-qPCR: suitable

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

probe-based

shipped in

wet ice

storage temp.

−20°C

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. This convenient 2X master 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
  • JumpStart™Taq 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.
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

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/vialSDS
  • M8787Magnesium chloride solution, PCR Reagent, 25 mM MgCI2 solution for PCR 25 mMSDS
  • P219210X PCR Buffer, Optimized for routine PCR with MgCl2 included 100 XSDS

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Pictograms

Exclamation markEnvironment
GHS07,GHS09

Signal Word

Warning

Hazard Statements

H315,H317,H319,H410

Hazard Classifications

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

Storage Class Code

12 - Non Combustible Liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Validation of Quantitative PCR Assays
Lovatt, A., et al.
BioPharm., 22-32 (2002)
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
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

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