Bio-Rad Laboratories, a multinational manufacturer of life science research and clinical diagnostic products, has introduced its iScript RT-qPCR Sample Preparation Reagent for the rapid isolation of total RNA, which enables reverse transcription and realtime PCR to be performed directly from cell lysates.
The company says scientists can use the protocol to remove genomic DNA and stabilise RNA in as little as five minutes.
"We designed iScript RT-qPCR Sample Preparation Reagent to meet the needs of researchers performing high-throughput gene expression analysis, RNAi and microarray validation," said Viresh Patel, senior product manager, PCR Reagents at Bio-Rad Laboratories.
The company says conventional RNA purification methods, such as organic extraction or column-based methods that include salting out or the use of magnetic particles, require numerous steps to purify RNA without contaminating genomic DNA and can take at least 30 minutes to complete. The iScript RT-qPCR Sample Preparation Reagent employs selective lysis of the cell membrane while leaving the nuclear membrane intact to achieve DNA-free RNA within five to 10 minutes.
"This reagent provides a more affordable option for researchers looking to complete sensitive, high-throughput gene expression analysis," added Patel.
Upon exposure of cells to iScript RT-qPCR Sample Prep Reagent, cell disruption and lysis occurs in approximately 30 seconds with the aid of mild vortexing. The reagent also stabilises RNA and effectively inhibits intracellular ribonucleases (RNases). A short two-minute high-speed centrifugation yields total RNA that can be collected in the supernatant while the nuclei containing genomic DNA form a pellet and can be discarded. The resulting RNA preparation is ready for use, without further treatment, in subsequent reverse transcription and PCR/qPCR.
RNA isolated with the iScript RT-qPCR Sample Preparation Reagent is stable for up to six months when stored at -20°C.
