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ReadyScript® cDNA Synthesis Mix

Complete reagent for first strand cDNA synthesis for RT-qPCR

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Synonym(s):
cDNA Synthesis Mix

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

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This Item
P460011483188001FPCR-RO
ReadyScript® cDNA Synthesis Mix Complete reagent for first strand cDNA synthesis for RT-qPCR

RDRT

ReadyScript® cDNA Synthesis Mix

ReadyMix™ Taq PCR Reaction Mix with MgCl2

P4600

ReadyMix Taq PCR Reaction Mix

FastStart™ PCR Master sufficient for ≤100 reactions (04710436001), sufficient for ≤400 reactions (04710444001), sufficient for ≤2,000 reactions (04710452001)

FPCR-RO

FastStart PCR Master

usage

sufficient for 100 reactions

usage

sufficient for 100 reactions

usage

sufficient for 30 reactions (including 5 control reactions)

usage

sufficient for ≤2,000 reactions (04710452001), sufficient for ≤100 reactions (04710436001), sufficient for ≤400 reactions (04710444001)

feature

dNTPs included

feature

dNTPs included, hotstart: no

feature

dNTPs included, hotstart: no

feature

-

color

colorless

color

colorless

color

-

color

-

input

purified RNA

input

purified DNA

input

purified RNA

input

-

shipped in

dry ice

shipped in

wet ice

shipped in

-

shipped in

-

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

10 - Combustible liquids

wgk_germany

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable


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Articles

One approach to the analysis of gene expression is to measure the concentration of mRNA of a gene. There are several challenges to such analyses, such as the differences in half life between different transcripts, the temporal patterns of transcription and the lack of correlation between mRNA and protein.

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

Our SYBR Green qPCR Protocol is a method designed to detect accurate quantification of gene expression and RT-PCR reactions

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.

RT-qPCR detects specific targets with applications in gene expression and pathogen detection.

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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