New Kits Available for RNA Isolation from FFPET Samples and for cDNA Preamplification

By LabMedica International staff writers
Posted on 13 Nov 2012

Image: The High Pure FFPET RNA Isolation Kits (Photo courtesy of Roche).

A new kit will allow genomics researchers to isolate highly purified samples of RNA from FFPET (formalin-fixed, paraffin-embedded tissue) samples for analysis by microarray or for the generation of DNA for PCR studies.

The new High Pure FFPET RNA Isolation Kit was recently placed onto the biotech market by Roche (Basel, Switzerland). This kit was designed for the isolation and purification of total RNA from 5 to 10 micrometer thick sections from FFPET. The isolated RNA is suitable for qRT-PCR (real-time PCR) and microarray analysis.

An investigator using the High Pure FFPET RNA Isolation Kit will require approximately 2.5 hours including the deparaffinization procedure to prepare an RNA sample. The deparaffinized tissue is lysed with special lysis buffers and incubated with Proteinase K. In the presence of chaotropic salts, the RNA is specifically bound to the glass fiber of the kit's High Pure Filter Tube. Bound RNA is then incubated with DNAse and purified in a series of rapid wash-and-spin steps and then eluted from the column. The RNA is recovered without contamination in an elution volume of 25-50 microliters.

The isolated RNA can be analyzed directly, or it may be transcribed into cDNA and subsequently preamplified with Roche's new RealTime Ready cDNA Pre-Amp Master Kit. This kit enables preamplification of minute amounts of cDNA with customized primer pools that correspond to primers that can be applied in subsequent qPCR analysis.

“Our new products provide highly convenient and flexible solutions to researchers who are analyzing gene expression in FFPET samples, while ensuring excellent result quality,” said Gerd Haberhausen, business head of qPCR and sample preparation systems at Roche. “By offering customized, aligned primer sets alongside our new RNA isolation kit, we are able to provide a complete and reliable workflow to scientists investigating the mRNA expression profiles of complex biological pathways.”

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