• Optimized for Real Time PCR with SYBR^® Green I
• Easy Preparation
• Extremely High Specificity

Description

SYBR^® Premix Ex Taq™ II (Perfect Real Time) is real-time PCR specific reagent by intercalater method using SYBR^® Green I^*1. This is a double concentration premix type reagents, which contains SYBR^® Green I ; its concentration is adjusted to real-time monitoring. This makes preparation of reaction mixture easer.
In comparison to SYBR^® Premix Ex Taq™ (Perfect Real Time) (Cat.# RR041A), this product improved buffer system, which resulted in improving reaction specificity. More wide range and accurate measurement is possible by controlling non-specific amplification, which prevent measurement.
The combinations of this buffer and Hot Start PCR enzyme TaKaRa Ex Taq™ HS that uses anti-Taq-antibody made possible to perform highly reliable expresstion with real-time PCR analysis.

Applicable real time PCR instruments;

• Thermal Cycler Dice™ Real Time System (TaKaRa Cat.# TP800)
• ABI PRISM^® 7000/7700,^*2 Applied Biosystems 7300/7500 Real-Time PCR System (Applied Biosystems)
• LightCycler^® (Roche Diagnostics) ^*3

(Note) It is recommended to use previous product (Cat.# RR041A) for Smart Cycler^® System / Smart Cycler^® II System (Cepheid).

*1 SYBR^® Green I is provided under licensing agreement with Molecular Probes Inc. for internal research use only.
SYBR^® is a registered trademark of Molecular Probes Inc.
*2 ABI PRISM^® is a registered trademark of Applera Corporation.
*3 LightCycler^® is a registered trademark of Roche Diagnostics.

Components

(200 reactions×50 µl PCR)

SYBR® Premix Ex Taq™ II (Perfect Real Time) (2×conc) *1		1.0 ml×5
ROX™ Reference Dye (50×conc.) *2		200 µl
ROX™ Reference Dye II (50×conc.) *2		200 µl

*1 contains TaKaRa Ex Taq™ HS, 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 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. It is not required for use with LightCycler^® and Thermal Cycler Dice™ Real Time System.

ROX™ is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries.

Storage

Stable at 4°C for 6 months.
It should be protected from light, and careful attention should be made not to cause contamination.

* This product is shipped on dry ice. After delivered, store at 4°C protected from light.
Make the concentration uniform by gently inverting a tube before use.

* The storage at -80°C is most recommended for long term storage. Once thawed, store at 4°C and use up within 6 months.

Note:
The enzyme activity may be lowered when stored in a freezer set at -20°C. This is because some freezers do not strictly control the temperature, accordingly SYBR^® Premix Ex Taq™ II can be thawed in a freezer. The most secured storage is to keep the products at -80°C before use, and store SYBR^® Premix Ex Taq™ II at 4°C after thawed.

Note

TaKaRa Ex Taq™ HS used in this product is an enzyme for hot start PCR utilizing Taq antibody.
Initial denaturation step prior to PCR should be 95°C for 30 sec. It does not require the activation step of heating at 95°C for 10-15 min prior to the PCR.

Application

For Real time PCR quantification of DNA using SYBR^® Green I

Features

• Quick and accurate detection and quantification of target gene through real time PCR.


• Easy-to-use 2× premix reagent including SYBR^® Green I:
  Ready to perform real time PCR in the presence of a fluorescent intercalator. Just add PCR primers, template and dH₂O to start the reaction.


• High amplification efficiency and highly sensitive detection:
  This kit utilizes an enzyme for hot start, TaKaRa Ex Taq™ HS. As this enzyme-buffer system is optimized for real time PCR, this product offers highly effcient amplification and high sensitivite detection.
• More wide range and accurate measurement is possible by the improved buffer system, which controls non-specific amplification and realizes excellent reaction specificity.

PCR test

Consistent amplification results and SYBR^® Green I detection were observed in real time PCR using HL60 cDNA as a template (amplified fragment: 56, and 184 bp) and Applied Biosystems 7500 Real-Time PCR System as a real time PCR instrument.