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ReadyScript® cDNA Synthesis Mix

Complete reagent for first strand cDNA synthesis for RT-qPCR

Synonym: cDNA Synthesis Mix

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Properties

Related Categories Molecular Biology, PCR/Amplification, RT-PCR and RT-qPCR More...
color   colorless
shipped in   dry ice
storage temp.   −20°C

Description

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 Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 2
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

Reverse Transcription - PCR Technologies Guide

Use the table of contents on the right to navigate to other sections of A Technical Guide to PCR Technologies.
A Technical Guide to PCR Technologies
Keywords: Degradations, Gene expression, Nucleic acid hybridization, PAGE, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, Transcription

Protocols

Imprint RIP | uRNA Quantitation | MicroRNA primers | MicroRNA Reverse Transcription

The Imprint® RNA Immunoprecipitation Kit was used to co-purify human argonaute 2 (Ago2)-associated RNAs from HeLa cells, as idicated in the Technical Bulletin. To evaluate the different stringency op...
Carol Kreader
Principal R&D Scientist
Keywords: Buffers, Cell disruption, Immunoprecipitation, Polymerase chain reaction - quantitative, RNA immunoprecipitation, Western blot

Reverse Transcription Using ReadyScript cDNA Synthesis RDRT Protocol

Place components on ice. Mix, and then briefly centrifuge to collect contents at the bottom of the tube
Keywords: Amplification, Digestions, Gene expression, Polymerase chain reaction, Polymerase chain reaction - quantitative, Precipitation, Purification, RNA purification, Transcription

Standard Reverse Transcription Protocol (Two-step)

Reverse transcription (RT) is the process of converting RNA to cDNA using a reverse transcriptase enzyme and dNTPs.
Keywords: Amplification, Polymerase chain reaction, Polymerase chain reaction - quantitative, Transcription

Universal SYBR Green qPCR Protocol

Technology Overview Assay Considerations Methods of Quantification Equipment & Supplies PCR Mix Selection Guide Protocol Troubleshooting Materials References
Keywords: AGE, Amplification, Buffers, Degradations, Electrophoresis, Enzyme activity, Gas chromatography, Gel electrophoresis, Gene expression, Genetic, Melting, Microarray Analysis, Nucleic acid annealing, Nucleic acid denaturation, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymorphisms, Purification, Sample preparations, Size-exclusion chromatography, Solvents, Titrations

Related Content

PCR Selection Guide

We offer a wide variety of PCR enzymes, master mixes, and PCR protocols to meet your experimental needs for routine PCR, qPCR, or RT-PCR. Our PCR Selection Guide features various filters to sort by, ...
Keywords: Genomics, Polymerase chain reaction, Polymerase chain reaction - quantitative

RT-PCR | Reverse Transcription PCR

Reverse transcription polymerase chain reaction (RT-PCR) is a variation of standard PCR that involves the amplification of specific mRNA obtained from small samples. It eliminates the need for the te...
Keywords: Amplification, Cloning, Diagnostic, Gas chromatography, Molecular biology, Nucleic acid hybridization, PAGE, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, Transcription

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 o...
Keywords: Amplification, Buffers, Clinical, Cloning, Diagnostic, Diseases, Gas chromatography, Gene expression, Homogenization, Infectious Diseases, Nucleic acid annealing, Nucleic acid hybridization, PAGE, Polymerase chain reaction, Polymerase chain reaction - quantitative, Transcription

Peer-Reviewed Papers
15

References

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