GeneMATRIX Gram Plus & Yeast Genomic DNA Purification Kit is designed for the rapid purification of genomic DNA from a wide variety of gram positive bacterial physiological groups and from a wide variety of yeast strains. The mechanical lysis procedure used within protocol is a universal approach for efficient cell disruption, avoiding to use expensive lytic enzymes specific to a particulary group of organisms. Purified DNA is free of contaminants, such as: RNA, proteins, lipids, dyes, detergents, organic inhibitors of enzymatic reactions, buffers, salts, divalent cations, among others. 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. An optimized buffer is added to provide selective conditions for DNA binding during a brief centrifugation step, while contaminants pass through the GeneMATRIX membrane in the spin-column. Traces of contaminants remaining on the membrane are efficiently removed in two wash steps. High-quality cellular DNA is then eluted in low salt buffer, e.g.: Tris-HCl, TE or water. Isolated DNA is ready for downstream applications without the need for ethanol precipitation.
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