The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Patent No.:

US 9777266 B1

PDF
Full Text
Expired
Date of Patent:
Oct. 03, 2017

Filed:

Sep. 02, 2016
Applicant:

The Johns Hopkins University, Baltimore, MD (US);

Inventors:

Elad Firnberg, Baltimore, MD (US);

Marc Alan Ostermeier, Baltimore, MD (US);

Assignee:

THE JOHNS HOPKINS UNIVERSITY, Baltimore, MD (US);

Attorneys:
Primary Examiner:
Int. Cl.
CPC ...
C12P 19/34 (2006.01); C12N 15/10 (2006.01); C07H 21/04 (2006.01);
U.S. Cl.
CPC ...
C12N 15/1031 (2013.01); C07H 21/04 (2013.01); C12N 15/102 (2013.01); C12N 15/1058 (2013.01); C12P 19/34 (2013.01);
Abstract

Methods for the creation of one or more user-defined mutations that can be located anywhere in a target sequence, such as in a gene are disclosed. These methods can be used on a single-stranded or double-stranded template and, for a single-stranded template, generally include: (a) conducting a first amplification reaction in the presence of a thermostable DNA polymerase and a thermostable DNA ligase to synthesize a mutagenized strand of DNA comprising at least one mutagenic oligonucleotide relative to a complementary single-stranded template DNA comprising a target nucleic acid molecule in a circular DNA vector; (b) conducting a second amplification reaction in the presence of a thermostable DNA polymerase and a thermostable DNA ligase to synthesize a complementary mutant strand of DNA; and (c) degrading the template DNA and non-covalently closed circular nucleic acid molecules to obtain a mutation-containing double-stranded DNA product.


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