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Sigma-Aldrich

Quantitative RT-PCR ReadyMix™

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

Synonyme(s) :

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

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About This Item

Code UNSPSC :
41106300
Nomenclature NACRES :
NA.55

Niveau de qualité

300

Utilisation

sufficient for 250 reactions

Caractéristiques

dNTPs included
hotstart

Technique(s)

RT-qPCR: suitable

Couleur

colorless

Entrée

purified RNA

Compatibilité

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

Méthode de détection

probe-based

Conditions d'expédition

wet ice

Température de stockage

−20°C

Description générale

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).

Caractéristiques et avantages

  • 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

Conditionnement

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

Informations légales

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

Composants de kit seuls

Réf. du produit
Description
  • Probe Based qRT-PCR ReadyMix 2 X
  • Moloney Murine Leukemia Viral Reverse Transcriptase (M-MLV RT) 5000 U

Composants de kit également disponibles séparément

Réf. du produit
Description
FDS
  • P219210X PCR Buffer, Optimized for routine PCR with MgCl2 included 1.5 mL/vialFDS
  • M8787Magnesium chloride solution, PCR Reagent, 25 mM MgCI2 solution for PCR 25 mMFDS
  • P219210X PCR Buffer, Optimized for routine PCR with MgCl2 included 100 XFDS

Produit(s) apparenté(s)

Réf. du produit
Description
Tarif

MRN70

GenElute™ mRNA Miniprep Kit, sufficient for 70 purifications

DMN70

GenElute™ Direct mRNA Miniprep Kits, sufficient for 70 purifications

RTN350

Kit Miniprep pour ARN total de mammifère GenElute™, sufficient for 350 purifications

T3934

Réactif TRI®, LS, For processing fluid samples such as cell suspensions, CSF, and amniotic fluid.

RTN70

Kit Miniprep pour ARN total de mammifère GenElute™, sufficient for 70 purifications

T9424

Réactif TRI®, For processing tissues, cells cultured in monolayer or cell pellets

DMN10

GenElute™ Direct mRNA Miniprep Kits, sufficient for 10 purifications

T3809

Réactif TRI®, BD, For processing whole blood, plasma, or serum.

RTN10

Kit Miniprep pour ARN total de mammifère GenElute™, sufficient for 10 purifications

AMPD1

DNase I, Amplification Grade

MRN10

GenElute™ mRNA Miniprep Kit, sufficient for 10 purifications

Pictogrammes

Exclamation markEnvironment
GHS07,GHS09

Mention d'avertissement

Warning

Mentions de danger

H315,H317,H319,H410

Classification des risques

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

Code de la classe de stockage

12 - Non Combustible Liquids

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable

Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.

Consulter la Bibliothèque de documents

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

Articles

Quantitative RT-PCR

Sigma’s 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.

Examination of Efficacy, Specificity, and Efficiency of an shRNA Lentiviral Library

The introduction of small interfering RNAs (siRNAs) into cultured cells provides a fast and efficient means of knocking down gene expression and has allowed siRNAs to quickly become a ubiquitous tool in molecular biology.

Examination of Efficacy, Specificity and Efficiency of an shRNA Lentiviral Library

Introduction of small interfering RNAs (siRNAs) into cultured cells provides a fast and efficient means of knocking down gene expression and has allowed siRNAs to quickly become a ubiquitous tool in molecular biology.

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

Protocoles

Reverse Transcription Protocol (One-step Probe Detection)

Reverse transcription (RT) is the process of converting RNA to cDNA using a reverse transcriptase enzyme and dNTPs.

Coronavirus qPCR Design Case Study & Related Products to Support SARS-CoV-2 Research

Coronavirus qPCR Primer and probe sets for the detection of SARS-CoV-2 (Corona Virus), with additional real-time RT-PCR, RT-qPCR and supporting reagents available.

Contenu apparenté

RT-qPCR – Quantitative Reverse Transcription PCR

RT-qPCR, or quantitative reverse transcription PCR, combines the effects of reverse transcription and quantitative PCR or real-time PCR to amplify and detect specific targets. RT-qPCR has a variety of applications including quantifying gene expression levels, validating RNA interference (RNAi), and detecting pathogens such as viruses.

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