SunScript™ One Step RT-PCR Kit
SunScript™ One Step RT-PCR Kit is an easy and reliable system designed for fast RT-PCR (real time) reactions based on detection of double-stranded DNA-binding dyes. One key advantage of the kit is the elevated temperature for performing the RT step thus allowing higher specificity of the reaction, and a higher likelihood in obtaining reaction products for difficult targets. Moreover, the excellent kinetic performance of the SunScript™ Reverse Transcriptase allows for speed optimization of the protocol, resulting in reaction times as low as 5 minutes for complete cDNA synthesis.
Features and Benefits:
RT step completed in as little as 5 minutes
RT reaction temperatures of at least 75°C
Outperforms one step RT-qPCR competitors
Easy detection of low abundance gene products
Broad dynamic range of quantification
Accurate quantification for high quality standard curves
Compatible with all qPCR machines (Sybr Green quantification)
The system uses an enzyme master mix containing:
SunScript™ Reverse Transcriptase RNaseH+, a high quality HotStart Taq Polymerase,
2X buffer containing dNTPs, salts, stabilizers and the double stranded DNA-binding dye
SunScript™ Reverse Transcriptase RNaseH+ is an engineered version of the well characterized HIV-1 RT with increased thermostability. This enables to perform the reverse transcription step at temperatures up to 75°C if needed (the recommended temperature for the RT reaction is 60°C for most samples depending on the melting temperature of the primers). These features make this kit the best choice for amplifying a broad range of mRNAs, also with complicated secondary structures or high GC content.
The reverse transcription reaction can be effectively performed while Taq Polymerase remains inactive during this step due to blocking antibodies
The inactivation/activation step at 95°C provides an automatic hot start for the Taq Polymerase to amplify the reverse transcribed cDNA
Intercalated SYBR Green I can be detected using common SYBR Green I settings (excitation wavelength ~450 nm, emission wavelength~ 520 nm)
The reaction mixes contain proprietary formulations which have been optimized for both the reverse transcription and the qPCR reaction, including dNTPs, MgCl2 and stabilizers resulting in a final concentration of 250 nM each dNTP and 3.5 mM MgCl2 in the final reaction.
The final recommended reaction volumen is 20 µl, suitable for all plate-based qPCR instruments (e.g. Bio-Rad CFX series Real-Time PCR Detection Systems, Roche LightCycler™ 480, etc.).
Shipping and Storage:
This product is shipped on dry ice. Upon receipt, it should be stored immediately at -20°C in a non-frost-free (constant temperature) freezer. If stored correctly, the product can be kept for at least three months after shipping without displaying any reduction in performance. For longer periods of time, store the kit at -80°C. Avoid frequent freeze-thaw cycles. Avoid longer exposure of kit contents to light.
Each batch of SunScript™ One Step RT-qPCR Kit and individual components are tested against predetermined specifications to ensure consistent product quality.
SunScript™ One Step RT-qPCR Kit is intended for molecular biology use only and in vitro use only. This product is not intended for diagnosis, prevention or treatment of a disease in human beings or animals.
Safety data sheets are available for download. This material may contain substances or activities that are harmful to human health. It should not be ingested, inhaled, or brought into contact with skin, and handled with appropriate care in accordance with the principles of good laboratory practices.
1 x 45 µl
2x Reaction Buffer
1 x 1000 µl
1 x 1000 µl