Applied Biosystems has announced a line of reagents that provide researchers with new ways of streamlining the process of performing quantitative reverse transcription PCR in RNA-based research
These reagents consist of new formulations and product-line improvements which are expected to provide researchers with a range of workflow solutions to simplify the processing of RNA in research applications such as gene expression and pathogen detection.
Real-time PCR, which is widely used in academic institutions as well as the biotechnology, pharmaceutical and medical research industries, is a laboratory method used to detect and determine simultaneously the amount of nucleic acids present in samples.
In RNA-based research applications, the use of reverse transcription is critical to the real-time PCR workflow.
By utilising reverse transcription reagents, RNA can be transcribed into single-strand cDNA, making PCR analysis possible.
This process helps researchers to assess more precisely how changes in the amount of RNA contribute to the development of disease or how they relate to biological processes in cells.
The following new products expand Applied Biosystems's portfolio of reverse transcription PCR reagents:.
High Capacity RNA-to-cDNA Master Mix for first strand cDNA synthesis in high throughput applications.
High Capacity RNA-to-cDNA Kit for first strand cDNA synthesis in low to medium throughput applications.
TaqMan RNA-to-CT 2-Step Kit and Power SYBR Green RNA-to-CT 2-Step Kit for screening a large number of targets from the same sample.
TaqMan RNA-to-CT 1-Step Kit (to be released April 2008) and Power SYBR Green RNA-to-CT 1-Step Kit for one-step workflows with reduced hands-on processing.
These new reverse transcription reagents are specifically validated and optimised for use with Applied Biosystems's assays and its industry leading line of real-time PCR instrument systems.
The two-step reverse transcription PCR kits offer flexibility when a researcher is screening for multiple targets or prefers to archive cDNA for gene quantitation.
With the one-step real-time reverse transcription PCR kits, all reactions from reverse transcription to quantitative PCR occur in a single tube.
One-step reagent kits are most effective when detecting a single target from many samples or when a streamlined process with minimal chance of cross-contamination is desired.
"Applied Biosystems continues to expand its portfolio of PCR-related chemistries better tailored to our customers' workflows," said Peter Dansky, president of the functional analysis division of Applied Biosystems.
"Integrating these specialised reagents with our industry-leading real-time PCR instrument systems, comprehensive line of assays and software analysis tools enables our customers to perform quantitative reverse transcription PCR experiments that accelerate their research in gene expression and pathogen detection".
Scientists at Belgium's Veterinary and Agrochemical Research Centre are using the Power SYBR Green RNA-to-CT 1-Step Kit as part of a real-time PCR workflow to study the immune reaction of chickens after highly pathogenic avian influenza infection.
In influenza infection, RNA initiates the viral replication.
Real-time reverse transcription PCR allows this team of scientists to examine viral avian influenza RNA more closely and to generate host gene expression data.
These data inform research projects aimed at protecting human health and certifying the safety of food products.
"The use of real-time PCR is vital to the research conducted in our laboratory," said Steven Van Borm, a senior scientist at the Veterinary and Agrochemical Research Centre.
"The more we can do to increase throughput cost-effectively, without compromising the quality of results, the better equipped we will be to generate reliable data.
"The simplified workflow of the Power SYBR Green RNA-to-CT 1-Step Kit has allowed us to successfully save optimising assays with fewer steps while maintaining a high level of quality which helps us learn more about avian influenza viruses".
