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Key Documents

RDRT

Sigma-Aldrich

ReadyScript® cDNA Synthesis Mix

Complete reagent for first strand cDNA synthesis for RT-qPCR

Synonyme(s) :

cDNA Synthesis Mix

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About This Item

Code UNSPSC :
12352200
Nomenclature NACRES :
NA.55

Utilisation

sufficient for 100 reactions
sufficient for 25 reactions
sufficient for 500 reactions

Caractéristiques

dNTPs included
hotstart: no

Technique(s)

RT-PCR: suitable

Couleur

colorless

Entrée

purified RNA

Conditions d'expédition

dry ice

Température de stockage

−20°C

Description générale

ReadyScript® cDNA Synthesis Mix is a sensitive and easy-to-use solution for two-step RT-PCR. This 5X concentrated mix provides all necessary components (except RNA template) for first-strand synthesis including: buffer, dNTPs, MgCl2, primers, RNase inhibitor protein, ReadyScript reverse transcriptase and stabilizers. ReadyScript is a RNase H(+) derivative of MMLV reverse transcriptase, optimized for reliable cDNA synthesis over a wide dynamic range of input RNA. The unique blend of oligo (dT) and random primers in the ReadyScript cDNA Synthesis Mix works exceptionally well with a wide variety of targets. It is optimized for the production of targets < 1kb in length. ReadyScript cDNA Synthesis Mix produces excellent results in both real-time and conventional RT-PCR.

Application

ReadyScript® cDNA Synthesis Mix has been used for complementary DNA (cDNA) synthesis from total/mRNA in 2-step reverse transcription polymerase chain reaction (RT-PCR) and quantitative reverse transcription polymerase chain reaction (RT-qPCR) assays for gene expression studies.

Caractéristiques et avantages

  • Specific ratio of randomers and oligo (dT) primers in the mastermix ensures optimal representation of all transcript sequences in the cDNA product
  • High reproducibility: Premixed master mix eliminates variability inherent in other kits requiring mixing of components
  • RNase H(+) Moloney murine leukemia virus (MMLV) reverse transcriptase ensures residual RNA does not interfere with PCR amplification
  • Reverse transcribes both rare and abundant genes with a wide input range of template
  • Convenient 5X master mix formula provides for quick and easy reaction set up; just add RNA template
  • Convenient 5X master mix formula provides for quick and easy reaction set up; just add RNA template
  • Includes proprietary blend of randomers and oligodT primers and an RNAse H+ derivative of Moloney murine leukemia virus (MMLV) reverse transcriptase
  • Allows for unbiased, accurate conversion of RNA to cDNA in just 40 minutes
  • Can use up to 1 μg of RNA in a single 20 μL reaction; reactions can also be scaled up
  • Highly stable; can be stored at -20°C for up to 1 year; 4°C for up to one month

Composants

Supplied as a 5X reaction buffer containing optimized concentrations of MgCl2, dNTPs (dATP, dCTP, dGTP, dTTP), recombinant RNase inhibitor protein, ReadyScript reverse transcriptase, random primers, oligo(dT) primer and stabilizers.

Autres remarques

Stable for 1 year when stored in a constant temperature freezer at -20 °C. To extend product′s shelf-life, store the kit at -70 °C. Readyscript cDNA Synthesis Mix showed no loss in functional performance after 20 cycles of freezing on dry ice and thawing on ice. However, we recommend keeping the freeze-thaw cycles to a minimum.

Informations légales

ReadyScript is a registered trademark of Merck KGaA, Darmstadt, Germany

Code de la classe de stockage

10 - Combustible liquids

Classe de danger pour l'eau (WGK)

WGK 2

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable


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Consulter la Bibliothèque de documents

Effect of prolonged water stress on essential oil content, compositions and gene expression patterns of mono- and sesquiterpene synthesis in two oregano (Origanum vulgare L.) subspecies.
Morshedloo MR
Plant Physiology and Biochemistry, 111, 119-128 (2017)
Takashi Kato et al.
Biochemical and biophysical research communications, 549, 83-90 (2021-03-06)
PTEN and p53 are highly mutated in many cancers. These two tumor suppressors have critical functions in the nucleus, such as DNA repair, cell cycle progression, and genome maintenance. However, the in vivo functional relationship of nuclear PTEN and p53 is
Extracellular vesicles-derived microRNAs expression as biomarkers for neurological radiation injury: Risk assessment for space exploration.
Gaines D, et al.
Life sciences in space research, 32, 54-62 (2022)
Sharada H Sharma et al.
European journal of pharmacology, 796, 20-31 (2016-12-19)
Targeting tumor metabolism by natural products is a novel approach and provides rationale for anti-cancer drug discovery. The present study aims to explore the impact of morin and/or esculetin on c-myc induced energy metabolism in 1,2-dimethylhydrazine (DMH) induced colon cancer
Water transport through the intestinal epithelial barrier under different osmotic conditions is dependent on LI-cadherin trans-interaction
Agnes Weth
Tissue barriers (2017)

Protocoles

Method for reverse transcription of RNA into DNA. Uses a premixed reagent that contains reverse transcriptase, dNTPs, primers, RNase inhibitor and buffer. Fast generation of cDNA.

Reverse transcription analyzes mRNA gene expression, facing challenges like transcript half-life differences and temporal patterns.

Reverse transcription analyzes mRNA gene expression, facing challenges like transcript half-life differences and temporal patterns.

Reverse transcription analyzes mRNA gene expression, facing challenges like transcript half-life differences and temporal patterns.

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Contenu apparenté

La réaction en chaîne par polymérase (PCR pour "Polymerase Chain Reaction") est une technique d'amplification des molécules d'acide nucléique qui est couramment employée dans de nombreuses applications, incluant la RT-PCR, la PCR à démarrage à chaud, la PCR en point final, et bien d'autres.

Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.

Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.

La réaction en chaîne par polymérase (PCR pour "Polymerase Chain Reaction") est une technique d'amplification des molécules d'acide nucléique qui est couramment employée dans de nombreuses applications, incluant la RT-PCR, la PCR à démarrage à chaud, la PCR en point final, et bien d'autres.

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