Principle

Isolation of total RNA in flexible 8-well strip format and for high throughput in approved 96-well format

NucleoSpin® 8/96 RNA for automated purification of total RNA from animal or human cell cultures and tissue combines the stringency of guanidine-isothiocyanate lysis with effective digestion of genomic DNA by rDNase treatment and the speed of silica-membrane technology. This takes the application to the next level for use with common liquid handling instruments. Ready-to-run protocols are available at MACHEREY-NAGEL and can be customized to fit your needs. NucleoSpin® 8/96 RNA means consistent RNA quality, reproducible yields of RNA, and high-throughput procedures free of cross-contamination (see Application data). Therefore, NucleoSpin® 8/96 RNA kits are powerful products for e.g. microarray analysis in respect to gene expression profiling, drug discovery, and molecular diagnostics (see Application data). The kits allow purification of total RNA from up to 107 cells or up to 30 mg tissue (centrifuge processing). The unique 8-well strip/96-well plate design of the NucleoSpin® 8/96 RNA systems allow the fully automated drying of the RNA binding module before loading of elution buffer and minimize spraying. With the NucleoSpin® 8/96 RNA method (see procedure), cells or tissue are lysed by incubation in a solution containing large amounts of chaotropic ions (cells can be grown and directly lysed in 96-well cell culture plates). This lysis buffer immediately inactivates RNases and creates appropriate binding conditions which favor adsorption of RNA to the silica membrane of the NucleoSpin® 8 RNA Binding Strip/96 RNA Binding Plate. Contaminating DNA, which is also bound to the silica membrane, is removed by rDNase which is directly applied onto the membrane during the preparation (rDNase is supplied with the kits). Due to the high specific activity of the rDNase a treatment of only 15 minutes at room temperature is sufficient for an efficient removal of contaminating DNA (see Application data). Salts, metabolites, and macromolecular cellular components are removed by simple washing steps with three different buffers. Pure RNA is finally eluted under low ionic strength conditions with RNase-free water (pH 8.0–8.5).

Features