Applied Biosystems advances cell-based gene expression research

Applied Biosystems, an Applera Corporation business, today announced a new family of real-time PCR reagent kits that enable researchers to perform cell-based gene expression research more rapidly. The TaqMan Cells-to-CT™ Kit solutions are chemical reagents that are designed to reduce the amount of time that is typically necessary to prepare and analyse RNA from cultured cells using real-time PCR, in applications such as siRNA-mediated gene silencing and microRNA analysis.

This family of kits enables expression analysis to be performed directly from cultured mammalian cells derived from a variety of tissue sources, including epithelial cells and stem cells. Integrating gold-standard TaqMan chemistry, these kits are part of a complete workflow that has been designed with all the necessary reagents for performing expression analysis and are validated for use with Applied Biosystems’ TaqMan Assays and Arrays. These new, streamlined workflows eliminate the time-consuming step of RNA purification and reduce the traditionally labour-intensive process of preparing samples to under 10 minutes, compared to 30-60 minutes when using traditional methods.

These reagents, which are manufactured and validated by Applied Biosystems, are optimised for use with the company’s industry-leading real-time PCR instrument systems. The TaqMan Cells-to-CT kits expand Applied Biosystems’ portfolio of real-time PCR workflows. They include:

• TaqMan Gene Expression Cells-to-CT Kit for gene expression analysis from mRNA
• TaqMan PreAmp Cells-to-CT Kit for pre-amplification of selected targets when using precious or limited samples
• TaqMan MicroRNA Cells-to-CT Kit for microRNA expression analysis
• TaqMan Fast Cells-to-CT Kit (expected to be released May 2008) for gene expression analysis with Fast Real-Time PCR.

“The development of these new reagents demonstrates Applied Biosystems’ continued strategic focus on bringing high-quality, application-specific solutions for real-time PCR to market,” said Peter Dansky, president of Applied Biosystems’ functional analysis division. “Our customers are continuing to derive new applications for real-time PCR, and we intend to continue to play a leading role in providing them with differentiated solutions that are tailored to their specific research workflows.”
Real-time PCR is a laboratory method that is widely used in academic and industry laboratories to simultaneously detect and determine the amount of nucleic acids present in samples. Research data generated using these reagents can help researchers to assess precisely how changes in the amounts of RNA in these cells may contribute to the development of disease, as well as how they relate to biological processes.

For instance, Reproductive Medicine Associates of New Jersey, one of the world's largest and most experienced centres for infertility treatment, uses real-time PCR in its research, which focuses on developing new diagnostics and therapeutics for in vitro fertilisation. Scientists at the centre have developed an innovative procedure for screening all 46 human chromosomes to identify abnormalities in embryos, and have begun to determine when stem cells differentiate during pre-implantation embryonic development. They are using the TaqMan Gene Expression Cells-to-CT Kit for several projects including the characterisation of single cells in three-day old embryos. This particular kit allows them to rapidly process RNA and directly analyse it on TaqMan Arrays without purification. This is a new, time-saving workflow that is uniquely enabled by the kit. It also eliminates the loss of RNA, which is particularly important for downstream gene expression analysis of single cells.

"The use of real-time PCR is critical to our ability to perform expression analysis on stem cells," said Dr Nathan Treff, PhD, director of molecular biology at Reproductive Medicine Associates of New Jersey. "Using the TaqMan Gene Expression Cells-to-CT Kit and TaqMan Arrays, we have been able to bring efficiencies to our real-time PCR workflow, while increasing the throughput of our lab 10-fold."

The complete family of TaqMan Cells-to-CT Kit solutions has been extensively tested and validated to work with Applied Biosystems’ line of TaqMan MicroRNA Assays, and TaqMan Gene Expression Assays and TaqMan Arrays.