US 12,351,796 B2
Methods for rapid separation and purification of DNA topological forms
Massa Shoura, Stanford, CA (US); Stephen Levene, Redwood City, CA (US); and David Girata, Redwood City, CA (US)
Assigned to The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US); and BOARD OF REGENTS, THE UNIVERSITY OF TEXAS SYSTEM, Austin, TX (US)
Filed by The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US); and BOARD OF REGENTS, THE UNIVERSITY OF TEXAS SYSTEM, Austin, TX (US)
Filed on Mar. 7, 2024, as Appl. No. 18/598,993.
Application 18/598,993 is a continuation of application No. 18/098,615, filed on Jan. 18, 2023, granted, now 11,952,570.
Application 18/098,615 is a continuation of application No. PCT/US2021/042594, filed on Jul. 21, 2021.
Claims priority of provisional application 63/055,029, filed on Jul. 22, 2020.
Claims priority of provisional application 63/064,261, filed on Aug. 11, 2020.
Prior Publication US 2024/0218352 A1, Jul. 4, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/10 (2006.01)
CPC C12N 15/1006 (2013.01) [C12N 15/1003 (2013.01)] 20 Claims
 
1. A method for topology-dependent, rapid DNA purification of circular DNA, comprising:
combining a sample having DNA species with a chaotropic dense salt solution, wherein the DNA species comprises circular DNA in multiple topological states, wherein the combining is performed in the absence of intercalating dye and in the absence of protein-dependent digests;
stratifying the circular DNA in multiple topological states into different circular DNA topologies by ultracentrifugation of the DNA species in the chaotropic dense salt solution in the absence of intercalating dye to obtain stratified circular DNA topological states;
isolating at least one fraction of DNA of at least a first topological state in from the stratified circular DNA topological states;
removing excess salt from the isolated at least one fraction of DNA of the at least first topological state to generate a purified circular DNA of the at least first topological state.