Patent ID: 11873487
Assignee: UNIVERSITY OF MASSACHUSETTS
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method of enhancing homology directed repair efficiency of an isolated double-stranded nucleic acid donor sequence, the method comprising the steps of:
1) denaturing the double-stranded nucleic acid donor sequence to form two single-stranded nucleic acid sequences; and
2) renaturing the two single-stranded nucleic acid sequences to form the double-stranded nucleic acid donor sequence, thereby enhancing the homology directed repair efficiency of the double-stranded nucleic acid donor sequence,
wherein the double-stranded nucleic acid donor sequence has enhanced homology directed repair efficiency relative to a double-stranded nucleic acid donor sequence that has not been denatured and renatured;
wherein the double-stranded nucleic acid donor sequence is heat denatured or chemical denatured;
wherein, when the double-stranded nucleic acid donor sequence is heat denatured, the double-stranded nucleic acid donor sequence is denatured by incubating the double-stranded nucleic acid donor sequence at 80° C. to 100° C.; and
wherein the temperature is decreased after the denaturing step at a rate of about 0.1° C./second to about 2° C./second or the temperature is decreased immediately by placing the denatured donor sequence at about 4° C. to about 10° C. or by placing the denatured donor sequence on ice.