Cells are lysed by incubation in a solution containing chaotropic ions. This lysis buffer inactivates RNases creating appropriate binding conditions favouring adsorption of RNA to the silica membrane. After lysis, homogenisation and reduction of viscosity are achieved by filtration with NucleoSpin™ filter units. Contaminating DNA bound to the membrane is removed by a DNase I solution directly applied to the membrane during preparation. Optimal conditions for DNase are achieved by washing the membrane with a specific desalting buffer before treatment. Salts, metabolites and macromolecular cellular components are removed by simple washing with two different buffers. Total RNA is finally eluted with RNase-free water. The NucleoSpin™ RNA Plant kit features two alternative lysis buffers: in most cases, use of buffer RA1 is recommended for lysis due to its strong denaturing properties. The presence of peculiar metabolites in a variety of plant tissues or fungi may lead to solidification of the lysate, resulting in non-processible slurry. In such cases, the alternative buffer, RAP is the buffer of choice. For thorough homogenisation and reduction of lysate viscosity, NucleoSpin™ filter units are also provided. Kit components: NucleoSpin™ RNA columns with 2mL collecting tubes, 2mL collecting tubes, 1.5mL microcentrifuge tubes, NucleoSpin™ filters, buffers, RNase-free DNase I, DNase I reaction buffer, RNase-free water.
Silica membrane technology
Yield: 3μg to 70μg from 100μg plant material
Sample material: 1mg to 100mg tissue
Elution volume: 60μL
Fragment size: 200b to 20kb
Binding capacity: 200μg
Preparation time: 30min/6 preps
Format: mini spin column
DNase I included
NucleoSpin filter columns included for homogenisation of lysate and reduction of viscosity
Parallel purification of DNA possible by using the NucleoSpin RNA/DNA buffer set NucleoSpin™ RNA Plant is designed for isolation of DNA-free high quality RNA from a wide variety of plant and fungal samples.