Back to results
US 11,236,325 B2Granted
Methods for nucleic acid capture
AssigneeZymo Research Corporation
InventorsRyan Kemp, Jonathan A. Claypool, Marc E. Van Eden, Xi-Yu Jia
Life of this patent
Estimating the term…
Abstract
Solutions, reagents, and methods for nucleic acid purification. In certain aspects, cationic surfactant and, optionally, an anionic surfactant solutions are provided which can be used for phase separation and capture of nucleic acids, such as plasmid or genomic DNA, to a solid phase carrier, such as a mineral matrix.
Claims20
A method of isolating plasmid DNA by alkaline lysis, comprising:
- (a) resuspending cells comprising plasmid DNA in a first aqueous solution;
- (b) lysing the cells with a second solution, thereby generating a lysate comprising the plasmid DNA;
- (c) neutralizing the lysate with a third solution;
- (d1) capturing the plasmid DNA to a mineral matrix with a phase separation solution comprising domiphen bromide and in the presence of 0.1 M to 0.7 M LiCl salt;
- (d2) removing the phase separation solution from the mineral matrix and captured plasmid DNA;
- (e) washing the mineral matrix and captured plasmid DNA with a salt solution, thereby enhancing the retention of captured plasmid DNA;
- (f) washing the mineral matrix and captured plasmid DNA with an organic wash solution;
- and
- (g) eluting the plasmid DNA from the mineral matrix, thereby isolating the plasmid DNA.
6 more elements
Dependencies are inferred from the claim text.