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US 9,540,635 B2Granted
Buffer for one-step DNA extraction
AssigneeBio-Rad Laboratories, Inc.
InventorsBryony Ruegg, Ingrid Miller
Life of this patent
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Abstract
DNA is extracted from epithelial cells and other cells without cell walls, by use of a DNA extraction buffer that contains a C1-C4 alcohol, a cell lysis detergent, and a buffering agent maintained at a slightly basic pH. The cells are immersed in the buffer and gently agitated at ambient temperature, and DNA extraction and precipitation occur in thirty minutes or less, and often in five minutes or less.
Claims24
A process for extracting DNA from cells without cell walls, said process comprising:
- (a) immersing said cells in an aqueous solution comprising a C1-C4 alcohol at from about 50% to about 85% by weight, a cell lysis detergent at from about 0.05% to about 0.5% by weight, and a buffering agent maintaining a pH of from about 7.5 to about 9.0, and substantially no protease, substantially no reducing agent, and substantially no non-detergent, inorganic salt, at a temperature of from about 15° C. to about 35° C. for from about five seconds to about thirty minutes, to cause DNA to be liberated from said cells and to precipitate;
- and
- (b) recovering said DNA so precipitated from said aqueous solution.
A DNA extraction buffer comprising:
- (i) a C1-C4 alcohol at a concentration of from about 75% to about 90% by weight,
- (ii) a DNA extraction detergent at a concentration of from about 0.05% to about 0.5% by weight, and
- (iii) a buffering agent maintaining a pH of from about 7.5 to about 9.0,
- in aqueous solution containing substantially no protease, substantially no reducing agent, and substantially no non-detergent, inorganic salt.
1 more element
Dependencies are inferred from the claim text.