Description

SYBR^® Premix DimerEraser™ (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™ II (Perfect Real Time) (Cat. #RR081), this product is improved buffer system and is added with the original accessory protein, which result in improving reaction specificity.
This product has the effect in depression of primer dimer, which is especially important matter in SYBR Green Assay.
The product make possible to measure accurate quantitative analysis in a wide range concentration of template by suppression of non-specific amplification and highly detection of a lower amount of template.
We recommend a 3-step PCR method in the standard protocol of this product.

Applicable real time PCR instruments；

• ABI PRISM^® 7000,^*2 Applied Biosystems 7500 Real-Time PCR System, 7500 Fast Real-Time PCR System (Applied Biosystems)
• LightCycler^® (Roche Diagnostics)^*3
• Thermal Cycler Dice™ Real Time System (TAKARA BIO)
  
  This instrument is not available in the U.S. and Europe.
• Smart Cycler^® System/Smart Cycler^® II System (Cepheid)^*4

*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.
*4 Smart Cycler^® is a registered trademark of Roche Cepheid.

Components

(200 reactions×50 µl PCR)

SYBR Premix DimerEraser (Perfect Real Time) (2×conc.) *1	1 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. Refer to IX. Specifications for further details of TaKaRa Ex Taq™ HS.

*2

ROX™ Reference Dye/Dye II is used for normalization of intensity by background subtraction. For ABI PRISM® 7000, the use of ROX™ Reference Dye (50×) is recommended. For Applied Biosystems 7500 Real-Time PCR System and 7500 Fast Real-Time PCR System, the use of ROX™ Reference Dye II is recommended. It is not required for use with LightCycler®, Smart Cycler® System/Smart Cycler® II System and Thermal Cycler Dice™ Real Time System.

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. The storage at -20°C is not recommended. Once thawed, store at 4°C and use up within 6 months.

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

1. Quick & accurate detection and quantification of target gene through real time PCR.
2. Easy-to-use 2×premix reagent including SYBR^® Green I:
   Ready to perform real time PCR in the presence of a fluorescent intercalater. Just add PCR primers, template and dH₂O to start the reaction.
3. High amplification efficiency and highly sensitive detection:
   This product utilizes an enzyme for hot start, TaKaRa Ex Taq™ HS. As this enzymebuffer system is optimized for real time PCR, this product offers highly efficient amplification and high sensitive detection. Moreover, the addition of the accessory protein strongly suppress mis-priming of primers during PCR reaction and nonspecific amplification, such as primer-dimer.

Reagents and Instruments Required but Not Supplied in this product

1. Thermal Cycler for real time PCR
2. Reaction tube or plate for real time PCR
3. PCR primer
4. dH₂O
5. Micropippets and Micropippet tips (autoclaved prior to use)