Method for isolating and purifying nucleic acids

摘要

The present invention relates to a method for isolating and purifying nucleic acids, preferably comprising genomic DNA, from biological samples comprising the steps of lysing the sample using a lysis buffer comprising a source of anionic surfactant ions, optionally disintegrating the RNA present in the lysate, precipitating the surfactant ions from the lysate, and separating the nucleic acids from the precipitate and further contaminants by size-exclusion chromatography. The invention furthermore relates to a lysis buffer, a method of lysing cells and a kit for the isolation and purification of nucleic acids.

主权项

1. A method of lysing cells in a cell-containing biological sample comprising the steps of:
(1) mixing the sample with a lysis buffer, wherein the lysis buffer comprises a buffering substance, H2SO4 and a source of anionic surfactant ions, the buffer has a pH of 7.5 to 10, 8 to 9, or 8.5, and is essentially free of EDTA and Mg2+ ions, (2) incubating the mixture obtained in step (1) to obtain a lysate that comprises at least DNA, RNA and proteins, and is essentially free of a chelating or complexing agent, and (3) precipitating the surfactant ions out of the lysate.