NucleoSpin RNA II is recommended for isolation and purification of total RNA from cultured cells, tissue, bacteria, yeast, cell-free biological fluids, and reaction mixtures without the use of organic compounds (e.g., Trizol). This kit allows purification of up to 70 µg of highly pure, DNA-free total RNA. A reference RNA from 106 cultured HeLa cells is prepared following the standard protocol, resulting in 15 to 20 µg of total RNA. It is possible to detect transcripts from very small amounts of cells.
Total RNA prepared with NucleoSpin RNA II is suitable for applications such as reverse transcriptase-PCR (qRT-PCR), Northern blotting, primer extension, array technology, and RNase protection assays. For isolation of high-quality RNA, it is important to prevent degradation of the RNA and to eliminate genomic DNA. With the NucleoSpin RNA II method, cells are lysed in presence of chaotropic salts. The lysis buffer immediately inactivates RNases, which are present in virtually any biological material, and creates appropriate conditions for RNA to bind to the silica membrane. After lysis, homogenization and reduction of viscosity are achieved by filtration with NucleoSpin Filter units (shredders), which are provided with the kit. Residual genomic DNA is efficiently removed by on-column digestion with rDNase, which is also included in the kit.
