Extraction buffer and method for isolating high-quality RNA from cells exposed to metal chloride solutions and clay mineral suspensions
View Patent ↗An RNA extraction buffer, an RNA extraction method, and an RNA extraction kit are described which enable functional, rapid, efficient, and high-quality RNA isolation from samples containing high concentrations of aqueous metal cations, clays, silica, or silicate minerals.
1. A method for isolating RNA from bacterial cells exposed to one or more of clay mixtures, clay minerals, silicate minerals, phyllosilicates, and aqueous solutions including metal cations, comprising the steps of:
lysing at least one of said bacterial cells; and
extracting RNA separately from DNA with a mixture comprising a sodium phosphate buffer with a pH of about 4.5 to about 6.0, about 1.0 M to about 2.0 M NaCl, about 0.2% hexadecyltrimethylammonium bromide (CTAB), and a chelating agent.
2. The method of claim 1 , wherein the NaCl concentration is about 1.4 M.
3. The method of claim 1 , wherein the chelating agent is about 50 mM EDTA.
4. The method of claim 1 , wherein the extracting step further comprises using said buffer further including about 0.05% diethyl pyrocarbonate for about 12 hours, with diethyl pyrocarbonate subsequently inactivated by autoclaving, and about 1 mM 1,4-dithio-DL-threitol added immediately prior to use.