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Merck

RDRT

ReadyScript® cDNA Synthesis Mix

Complete reagent for first strand cDNA synthesis for RT-qPCR

Synonym(s):

cDNA Synthesis Mix

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About This Item

NACRES:
NA.55
UNSPSC Code:
12352200
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usage

sufficient for 100 reactions, sufficient for 25 reactions, sufficient for 500 reactions

feature

dNTPs included, hotstart: no

technique(s)

RT-PCR: suitable, cDNA synthesis: 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

  • 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

Storage Class

10 - Combustible liquids

wgk

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable


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Articles

Sigma Imprint RNA 免疫沉淀试剂盒用于从 HeLa 细胞中共纯化人 argonaute 2 (Ago2) 相关 RNA。MicroRNA通过Mysticq 试剂进行逆转录和定量。

分析基因表达的方法之一是测量基因的mRNA浓度。此类分析面临着若干挑战,例如不同转录本之间的半衰期不同、转录时间模式以及mRNA和蛋白质之间缺乏相关性。

Protocols

将RNA逆转录成DNA的方法。使用含有逆转录酶、dNTP、引物、RNase抑制剂和缓冲液的预混试剂。快速合成cDNA。

逆转录是一种旨在测量基因mrna浓度的基因表达分析。 这种分析存在多个挑战,例如不同转录本之间的半衰期差异、时间模式和缺乏相关蛋白。

我们的 SYBR Green qPCR 协议是专为检测基因表达和 RT-PCR 反应的精确定量而设计的方法。

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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