Patent ID: 11952570
Assignee: THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method for topology-dependent, rapid DNA purification of circular DNA greater than 50 kbp in size; the method comprising:
a) embedding cells in a polymer matrix;
b) dissolving the polymer matrix in a chaotropic dense salt solution to lyse the cells in situ, thereby providing a DNA sample comprising circular DNA greater than 50 kbp in size in multiple topological states in the absence of intercalating dye and in the absence of protein-dependent digests;
c) performing ultracentrifugation on the DNA sample in the chaotropic dense salt solution in the absence of intercalating dye;
d) isolating fractions of DNA from a gradient of the DNA sample created by the ultracentrifugation;
e) removing excess salt from the DNA fractions to generate a purified population of circular DNA greater than 50 kbp in size.