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RT-PCR

Maxime™ RT PreMix (Oligo (dT)15 Primer) 

Cat.No Capacity Inquire
25081 96 tubes Inquire
PRODUCT INFORMATION
Description

Dried & aliquoted RT PreMix Kit for simple and rapid cDNA synthesis from RNA of eukaryotic cells

  • • Excellent cDNA synthesis rate by OptiScriptTM RT System
  • • cDNA synthesis from a variety of RNAs by oligo dT15 primers
  • • Unnecessary for thermal pretreatment to solve secondary structure of RNA, possible for rapid reversibility
  • • Dried & aliquoted RT PreMix, All-in-one type
       - All components for reaction in dried pellet in a tube
       - Needed template and RNase free DW only for cDNA synthesis
  • • High stability and reproducibility
       - Excellent reproducibility with dried & aliquoted PreMix by ALHP system
       - Protection from oxidation and humidity by vacuum packaging

Maxime™ RT PreMix Kit (Oligo dT15 Primer) is a product that makes cDNA synthesis simple and fast from RNA of eukaryotic cells. This product is a product that can perform cDNA synthesis process in 1 step in a PCR tube using PCR machine unlike conventional 2 steps cDNA synthesis (RNA denaturation, cDNA synthesis). Maxime™ RT PreMix Kit (Oligo dT15 Primer) contains all components (Oligo dT15 Primer, dNTP mixture, Reaction buffer, etc.) required for RT reaction including OptiScript™ RT System, and it is contained in each PCR tube with dried pellet state by ALHP system. Because of this, it is possible to remarkably reduce the experimental error in pipetting between the experimenters, so the reproducibility is very high. With vacuum packaging method, it has excellent stability of activity due to the oxidation and moisture prevention of major components in the product. In addition, this product can synthesize cDNA easily with template RNA and RNase free DW addition. This product contains oligo dT15 primer, so it is easy to apply to eukaryotic RNA with poly (A) tail.

Applications
  • 01First strand cDNA synthesis
  • 02Construction of full length cDNA libraries
  • 03Real-time RT-PCR
  • 04RT-PCR
  • 05DNA labeling
  • 06Primer extension
Kit Contents
Contents 25081
Maxime™ RT PreMix Kit(Oligo dT15 Primer) 96 tubes
Manual 1 ea

[ Maxime™ RT PreMix Kit(Oligo dT15 Primer) ]

Component in 20 ml reaction

  • • OptiScript™ RT System
  • • Reaction Buffer
  • • RNase Inhibitor
  • • Primer – Oligo dT15 Primer
  • • Stabilizer
  • • dNTP Mixture
Technical Data

Comparison of cDNA synthesis performance with manual type

 

RNA from human was sequentially diluted from 1ug to one-third level, and cDNA was synthesized using Power cDNA Synthesis Kit, manual type cDNA synthesis kit, and Maxime™ RT PreMix Kit series. Then the efficiency of the synthesis was observed by PCR with Maxime PCR PreMix Kit (i-StarTaq ). As a result, the Maxime™ RT PreMix Kit (Oligo dT15 primer) showed equivalent performance to the existing manual type, Power cDNA Synthesis Kit, and the sensitivity was also comparable to Maxime™ RT PreMix Kit (Random primer).

Panel A, p53 gene (218bp) amplification; Panel B, Interleukin -10 gene(IL-10, 1.1kb) amplification
A, Power cDNA Synthesis Kit; B, Maxime RT PreMix Kit(oligo (dT)15 Primer); C, Maxime RT PreMix Kit(Random Primer)
Lane M, DNA Marker; lane N, no template control

Observation of quality stability


 

The RNA extracted from human cell (SNU-1) was serially diluted at intervals from 1ug to one-third level, and cDNA was synthesized using Maxime™ RT PreMix Kit (Oligo (dT)15 Primer). Then GAPDH gene (575 bp) was amplified by Maxime™ PCR PreMix Kit (i-StarTaq) to check stability according to storage period. As a result, it can be stabilized until 1 year. Maxime™ RT PreMix (Oligo (dT)15 Primer) is a product that keeps the performance of the product for 1 year under the freezing storage condition by preventing oxidation and incorporation of moisture by the vacuum compression manufacturing method.

TroubleShooting Guide
QThe product includes oligo dT 15mer size primer. Is there a product with oligo dT 20mer size primer?
AGenerally, oligo dT 15mer has been commonly used. So the product is provided with the oligo dT 15mer primer.
QHow long can be the synthesized cDNA stored?
AIt is recommended that the synthesized cDNA should be aliquoted with the amount used then stored at -70°C for 1 months.
QIn competitors products, there is RNA denaturation step. But iNtRON says that it doesn’t need a preheating step. Does not it have to proceed?
AIn order to differentiate from competitors, the product was designed to synthesize cDNA with 2 steps (45℃, 1hr / 95℃, 5min) without preheating to solve the secondary structure of RNA. Some customers said that they have achieved improved cDNA synthesis efficiency after doing preheating step. Therefore, it is up to you.
QWhat is the difference between Oligo dT primer and Random primer?
AOligo dT primers bind to the A-tail of mRNA to synthesize cDNA. Random primers randomly bind to rRNA, tRNA and total RNA as well as mRNA to synthesize cDNA. For mRNA target, Oligo dT primers are used commonly. But if A tailing is weak, it is recommended to use random primers.
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