RDRT

ReadyScript^® cDNA Synthesis Mix

Complete reagent for first strand cDNA synthesis for RT-qPCR

• Synonym(s): cDNA Synthesis Mix
• UNSPSC Code: 12352200
• NACRES: NA.55

Properties

• 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

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[1][2] in 2-step reverse transcription polymerase chain reaction (RT-PCR) and quantitative reverse transcription polymerase chain reaction (RT-qPCR)[3][4] assays for gene expression studies.[5][6]

Features and Benefits

• 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

• 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

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.

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.

Legal Information

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

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable