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One-Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time)

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The One-Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time) is a quantitative real-time PCR kit designed for SYBR Green I1-enabled product detection. Using this SYBR Real-Time PCR (qPCR) kit, RT-PCR can be performed in a single tube, minimizing pipetting steps and contamination risk. Reaction products are monitored in real time, eliminating the need for electrophoresis verification following PCR. The sensitivity of the enzymes offered in this SYBR real-time PCR kit enables detection of very small amounts of RNA, including RNA from viral, rare and precious samples.

 

The One Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time) takes advantage of PrimeScript RTase, an MMLV-based RNase H Minus reverse transcriptase that offers robust extension rates and synthesizes full-length cDNA quickly and easily. The kit also utilizes the TaKaRa Ex Taq HS high efficiency hot start PCR enzyme. Both kit enzymes are optimized for one-step RT-PCR via an optimized buffer. Combining the TaKaRa Bio RT-PCR technology with the following enzymes allows this one-step RT-PCR kit to generate high product yields. The premix formulation of the One Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time) also offers user convenience via a simplified protocol.


Technical Data:

Features

  • Quantitative Real Time PCR enables rapid and accurate analysis of RNA from viral, rare or precious samples.
  • TaKaRa Ex Taq HS is a high efficiency hot start PCR enzyme that can be used in RT-PCR.
  • The One-Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time) buffer system is optimized for quantitative real-time PCR, allowing efficient amplification and highly sensitive detection.
  • One Step SYBR RT-PCR Buffer IV is a 2X SYBR Green I premix; the premix formulation simplifies reaction mixture preparation.

Applications

  • Quantitative Real-Time PCR
  • SYBR Real-Time PCR (qPCR)
  • Detection and analysis of small RNA amounts

Components

RR086A: 100 reactions; 50 µL reaction system

2X One Step SYBR RT-PCR Buffer IV1 840 µL X 3
PrimeScript 1 step enzyme Mix II2 200 µL
RNase Free dH2O 1.25 mL X 2
ROX Reference Dye (50X conc.)3 100 µL
ROX Reference Dye II (50X conc.)3 100 µL
1 Includes dNTP Mixture, Mg2+ and SYBR Green I
2 Includes PrimeScript RTase, RNase Inhibitor and TaKaRa Ex Taq HS
3 ROX Reference Dye/Dye II are generally used for signal intensity normalization through background subtraction. For ABI PRISM 7000/7700/7900HT and Applied Biosystems 7300 Real-Time PCR System, the use of ROX Reference Dye (50X) is recommended. For Applied Biosystems 7500 Real-Time PCR System, the use of ROX Reference Dye II is recommended. The use of ROX Reference Dye or Dye II is optional; the Dye is not required for use with Thermal Cycler Dice Real Time System, SmartCycler or LightCycler real time instruments.ROX is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries.

Storage

–20°C. Protect 2X One Step SYBR RT-PCR Buffer IV from light.

References

1. Arita, M. et al. (2008) J. Gen. Virol. 89:2518-2530.

2. Murata, T. et al. (2011) J. Biol. Chem. 286:22007-22016.

Choosing a PrimeScript Kit for Real-Time RT-PCR

If you want to…. Use this….
Make cDNA using a master mix that includes RTase and primers PrimeScript RT Master Mix (Perfect Real Time)(Cat. # RR036A and RR036B)
Make cDNA using a kit that provides RTase and primers as separate components PrimeScript RT Reagent Kit (Perfect Real Time)(Cat. # RR037A and RR037B)
Avoid genomic DNA contamination during cDNA synthesis and RT-qPCR PrimeScript RT Reagent Kit with gDNA Eraser (Perfect Real Time) (Cat. # RR047A and RR047B)
Perform 1-step real-time RT-PCR with SYBR Green One-Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time) (Cat. # RR086A and RR086B)
Prepare cDNA for RT-qPCR directly from fewer than 1,000 cells CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 (Cat. # 3734)


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