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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

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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THIFICDNAROR252305893151001
technique(s)

RT-PCR: suitable

technique(s)

RT-PCR: suitable, RT-qPCR: suitable

technique(s)

PCR: suitable

technique(s)

RT-PCR: suitable, RT-qPCR: suitable, cDNA synthesis: suitable

usage

sufficient for 100 reactions

usage

sufficient for 100 reactions, sufficient for 200 reactions, sufficient for 50 reactions

usage

sufficient for 100 reactions, sufficient for 20 reactions

usage

sufficient for 100 reactions

feature

dNTPs included

feature

hotstart: no

feature

dNTPs included, hotstart: no

feature

dNTPs included: no, hotstart: no

color

colorless

color

-

color

red

color

-

input

purified RNA

input

purified RNA

input

purified DNA

input

purified RNA

shipped in

dry ice

shipped in

-

shipped in

wet ice

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 Sigma-Aldrich Co. LLC

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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