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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

application(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 Sigma-Aldrich Co. LLC

Storage Class Code

10 - Combustible liquids

WGK Germany

WGK 2

Flash Point F

Not applicable

Flash Point C

Not applicable

Certificate of Analysis

Certificate of Origin

Water transport through the intestinal epithelial barrier under different osmotic conditions is dependent on LI-cadherin trans-interaction
Agnes Weth
Tissue barriers (2017)
Effect of prolonged water stress on essential oil content, compositions and gene expression patterns of mono- and sesquiterpene synthesis in two oregano (Origanum vulgare L.) subspecies.
Morshedloo MR
Plant Physiology and Biochemistry, 111, 119-128 (2017)
Ashley Lui et al.
International journal of molecular sciences, 22(3) (2021-01-28)
Type 2 diabetes mellitus is a metabolic disorder defined by systemic insulin resistance. Insulin resistance in adipocytes, an important regulator of glucose metabolism, results in impaired glucose uptake. The trafficking protein, sortilin, regulates major glucose transporter 4 (Glut4) movement, thereby...
Agnes Weth et al.
Tissue barriers, 5(2), e1285390-e1285390 (2017-04-30)
In the intestine water has to be reabsorbed from the chymus across the intestinal epithelium. The osmolarity within the lumen is subjected to high variations meaning that water transport often has to take place against osmotic gradients. It has been...
Hui Li et al.
Toxicological sciences : an official journal of the Society of Toxicology, 164(2), 428-438 (2018-05-03)
Alcohol metabolism is a well-characterized biological process that is dominated by the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) families. Nonalcoholic steatohepatitis (NASH) is the advanced inflammatory stage of nonalcoholic fatty liver disease (NAFLD) and is known to alter the...

Protocols

Reverse Transcription Protocol

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.

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