This kit provides a method for an efficient and fast genomic DNA extraction from plant cells and tissues, or fungi.
It is known that plants contain quantities of different substances (polyssaccharides, polyphenols, etc) and that plants with the same or related genus can present enormous variabilities in their biochemical composition, for such reason, it becomes difficult to standardise on a single DNA extraction method for all plants.
To solve this problem and to be able to cover the biggest number of plants, DANAGENE uses a PVP solution that can bind the polyssacharides and polyphenols that are released by the cell lysis and has the capability of forming complexes with the nucleic acids to degrade them or to precipitate with them.
The process includes a sample homogenization in an extraction Buffer and the PVP solution. The lysis is completed with the incubation in a Lysis Buffer and RNase at 37°C for 30 minutes. Cell proteins and cell debris are removed by a protein removing Buffer that allows to leave the genomic DNA in solution. Finally, the genomic DNA is isolated by a precipitation with isopropanol.
This kit provides a method for an efficient and fast genomic DNA extraction from tissues of plants and fungi using MiniSpin columns.
The kit includes two optimized, alternative lysis buffers based on the established CTAB and SDS lysis methods. As plants are very heterogenous and contain a lot of different metabolites like polyphenols, polysaccharides, or acidic components, DANAGENE SPIN PLANT Kit offers two different lysis procedures for optimal processing of various samples.
In addition we also use a PVP solution that can bind the polyssacharides and polyphenols.
Procedure: Plant samples are first disrupted/homogenized and then lysed in a highly optimized buffer system, containing chaotropic salt, denaturing agent and detergents. A choice of two lysis buffers based on the established CTAB or SDS method are provided.
Crude lysate are cleared by centrifugation and the cleared lysate is then mixed with the Binding Buffer and processed through a MiniSpin column containing a silica membrane to which the plant genomic DNA binds.
Contaminants and impurities such a salts, metabolites and cellular components are removal by simple washing steps with two different buffers. High-quality purified plant genomic DNA is then eluted in a low Elution Buffer. The DNA is ready-to-use for a wide variety of applications.