Description

One Step SYBR^® PrimeScript™ RT-PCR Kit (Perfect Real Time) is designed for real time one-step RT-PCR using SYBR^® Green I^*1,*2 for detection. In this kit, RT-PCR can be performed all in a single tube subsequently, therefore the operation is simple, and it minimizes the risk of contamination. Also, amplified products are monitored in real time, so there is no need to verify them through electrophoresis after PCR. This kit is suitable for detection of tiny amount of RNA like RNA virus.
This kit uses PrimeScript™ RTase, which has excellent extendibility and can efficiently synthesize cDNA in short time period, and TaKaRa Ex Taq™ HS, high efficiency hot start PCR enzyme, and they are optimized for one step RT-PCR.

Applicable real time PCR instruments

• Thermal Cycler Dice™ Real Time System (TaKaRa Cat.#TP800)
• ABI PRISM^® 7000/7700/7900HT, 7300/7500 Real-Time PCR System (Applied Biosystems)
• LightCycler^® (Roche Diagnostics)
• Smart Cycler^® System^*3/Smart Cycler^® II System^*3 (Cepheid), and other

*1	SYBR® Green I is licensed by Molecular Probes Inc. for research reagents. SYBR® is a registered trademark of Molecular Probes Inc.
*2	Use One Step PrimeScript™ RT-PCR Kit (Perfect Real Time) for real time one-step RT-PCR using TaqMan® probe for detection. TaqMan® is a registered trademark of Roche Molecular Systems Inc.
*3	Smart Cycler® is a registered trademark of Cepheid.

This product is not available in the U.S.

Storage

-20°C
2× One Step SYBR^® RT-PCR Buffer III should be protected from light.

Kit Contents (for 100 reactions; 50 µl reaction system)

1.	2× One Step SYBR® RT-PCR Buffer III*1	840 µl × 3
2.	TaKaRa Ex Taq™ HS (5U / µl)	100 µl
3.	PrimeScript™ RT enzyme Mix II	100 µl
4.	RNase Free dH2O	1.25 ml × 2
5.	ROX Reference Dye (50×conc.)*2	100 µl
6.	ROX Reference Dye II (50×conc.)*2	100 µl

*1

Includes dNTP Mixture, Mg2+ and SYBR® Green I

*2

ROX™ Reference Dye/Dye II is used for normalization of intensity by background subtraction. For ABI PRISM® 7000/7700/7900HT and Applied Biosystems 7300 Real-Time PCR System, the use of ROX™ Reference Dye (50×) 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. It is not required for use with Thermal Cycler Dice™ Real Time System, Smart Cycler® or LightCycler® real time instruments.

Features

(1)	One Step RT-PCR made accurate and rapid analysis of RNA virus or small amount of RNA possible.
(2)	For PCR, enzymes for Hot Start TaKaRa Ex Taq™ HS, high efficiency hot start PCR enzyme, is used. Buffer system is optimized for Real-Time PCR, thus amplification is effective and high quality detection is possible. Moreover, One Step SYBR® RT-PCR Buffer III is a 2× concentration of premix with SYBR® Green I, so the preparation of reactants is simple.