GeneMATRIX Environmental DNA & RNA Isolation Kit is designed for rapid fractioning, isolation and purification of genomic DNA and total RNA starting from one single biological sample. Efficient lysis and cell disruption is warranted due to non-selective mechanical lysis supported by enzymatic treatment of the sample. This gives particulary good results when the bacteria are major target of the resercher.
Protocol outline: Sample material is mechanically disrupted, homogenized and lysed in the presence of special extraction buffer, which protect nucleic acids before degradation and reaction with humic substances. Bacterial or yeast cells are lysed in the presence of special cell wall disintegrating buffer aided by lysozyme (bacteria). The pretreated cell wall is then effectively destroyed by the mechanical friction of glass beads with optimally adjusted diameters. Further, Proteinase K digests cellular proteins, including stripping-off DNA of all bound proteins, among them nucleases. During the next steps, chloroform is added, and the homogenate is allowed to separate into a clear upper aqueous layer, an interphase, and a lower organic layer. Nucleic acids can be precipitated from the aqueous layer with isopropanol and separeted into DNA and RNA fraction using minicolumns and wash buffers. This kit is ideal for researchers who are interested in parallel studies of metagenomes and metatranscriptomes of a single sample.