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Real-Time qPCR & Reverse Transcription


Products >  Real-Time_qPCR_and_Reverse_Transcription >  Real-Time_qPCR_Probe_Detection >  Premix_Ex_Taq

Premix Ex Taq (Perfect Real Time)

Premix Ex Taq (Perfect Real Time) is a 2X RT-PCR kit specifically designed for fast and sensitive real-time PCR via either SYBR Green Real-Time PCR (qPCR) or probe-based qPCR assays. SYBR Green I Dye and probes for probe/5' nuclease based assays are not included in this RT-PCR kit. The Premix Ex Taq (Perfect RealTime) kit consists of Takara's high fidelity and high performance TaKaRa Ex Taq Hot Start polymerase and a real-time buffer that ensures superior specificity and increased amplification efficiency during RT-PCR (qPCR). Antibody-mediated hot start technology prevents nonspecific amplification due to mispriming and/or formation of primer-dimers during room temperature reaction assembly. Once the Taq antibody-polymerase complex is denatured during the first cycling step, the Premix Ex Taq polymerase can initiate DNA synthesis.

Two tubes of ROX Reference Dyes are supplied as separate components of this RT-PCR kit. ROX Dye is a convenient internal reference standard that may be used to normalize signal variances that occur as a consequence of non-PCR related fluorescence fluctuations across different wells or over time.

Premix Ex Taq (Perfect Real Time) provides superior specificity, performance and amplification yield during RT-PCR on all major real-time instruments.

Compatible Cyclers:

SmartCycler, Light Cycler, ABI PRISM 7000/7700/7900 HT, Applied Biosystems 7300/7500, iCycler, MJ Opticon, Stratagene MX 3000.

A dynamic range of 7–8 orders of magnitude using SYBR Green I and 10 orders of magnitude using probes can be expected with Premix Ex Taq (Perfect Real Time). Expected sensitivity is as low as 10 copies for both SYBR Green I and probe-based qPCR assays.

  At-A-Glance   Documents   Images & Data

Features

  • High specificity and increased amplification efficiency for qPCR
  • Sensitivity down to 10 copies for SYBR Green I or probe detection
  • Compatibility with 5' nuclease assays and with SYBR Green I Dye
  • Compatibility with many commonly used Real-Time PCR instruments

Applications

  • Real-Time PCR using probe-based qPCR assays or SYBR Green I Dye

Components

RR039A

Premix Ex Taq Mix (2X conc.)* 5 x 1 mL
ROX Reference Dye (50X conc.) 200 µL
ROX Reference Dye II (50X conc.) 200 µL

*contains TaKaRa Ex Taq HS DNA Polymerase, dNTP mix and Mg2+

RR039W

Premix Ex Taq Mix (2X conc.)* 1 x 25 mL
ROX Reference Dye (50X conc.) 1 mL

*contains TaKaRa Ex Taq HS DNA Polymerase, dNTP mix, and Mg2+

Storage

Product is stable at 4°C for 6 months following thaw. Protect from light and contamination. Long term storage should be at –20°C.

This product is shipped at –20°C. To ensure concentration uniformity, gently invert tubes prior to use.

PCR Performance

Consistent amplification results were confirmed by SYBR Green I detection of PCR products in real-time PCR using a 300 bp lambda DNA template and a Smart Cycler real-time PCR instrument. Inhibition of Ex Taq polymerase activity by the antibody was confirmed to be greater than 90% following 55°C incubation for 10 minutes.

Note: The ROX Reference Dye/Dye II is supplied for normalization of fluorescent signal intensities within wells when used with real-time PCR instruments which have this option. For ABI PRISM 7000/7700/7900HT and Applied Biosystems 7300 Real-Time PCR Systems, the use of ROX Reference Dye (50X) is recommended. For the Applied Biosystems 7500 Real-Time PCR System, use of ROX Reference Dye II is recommended. The use of ROX Reference Dye or Dye II is optional and not required when using Smart Cycler and LightCycler real-time instruments.

References

  1. Iwahara, K. et al. (2006) J. Clin. Microbiol. 44(1):172-176.
  2. Hayama E. et al. (2006) Pediatr. Res. 59(2):167-174.
  3. Jordan, E.N. & Durso, M.B. (2005) J. Mol. Diagnost. 7(5):575-581.

Additional Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


 
 
Products
Cat. # Product Contents Size Price License Units Select
RR039A Premix Ex Taq™ DNA Polymerase (Perfect Real Time) 200 Rxns $239.00 License Statements
RR039B Premix Ex Taq™ DNA Polymerase (Perfect Real Time) 400 Rxns $475.00 License Statements
RR039W Premix Ex Taq™ DNA Polymerase (Perfect Real Time), 25 mL 1,000 Rxns $953.00 License Statements
 

 

 
 

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