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RDRT

Sigma-Aldrich

ReadyScript® cDNA Synthesis Mix

Complete reagent for first strand cDNA synthesis for RT-qPCR

Synonym(s):

cDNA Synthesis Mix

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About This Item

UNSPSC Code:
12352200
NACRES:
NA.55

usage

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

feature

dNTPs included
hotstart: no

technique(s)

RT-PCR: suitable

color

colorless

input

purified RNA

shipped in

dry ice

storage temp.

−20°C

General description

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.

Features and Benefits

  • 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

Components

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.

Other Notes

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.

Legal Information

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

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

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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Articles

Sigma’s Imprint RNA Immunoprecipitation Kit was used to copurify human argonaute 2 (Ago2)-associated RNAs from HeLa cells. MicroRNAs reverse transcribed and quantitating using Mysticq reagents.

Protocols

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 is the analysis of gene expression to measure concentration mrna a gene. there are several challenges such analyses, as differences in halflife between different transcripts, temporal patterns and lack correlation protein.

Related Content

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.

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