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.

Date of Patent:
Jul. 20, 2021

Filed:

Apr. 12, 2019
Applicant:

Axiomx, Inc., Branford, CT (US);

Inventors:

Michael Weiner, Guilford, CT (US);

Margaret Kiss, Westbrook, CT (US);

Assignee:

AxioMx, Inc., Branford, CT (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C40B 50/06 (2006.01); C12N 15/10 (2006.01); C07K 16/00 (2006.01); C12N 15/90 (2006.01); C40B 30/04 (2006.01);
U.S. Cl.
CPC ...
C12N 15/1058 (2013.01); C07K 16/00 (2013.01); C12N 15/1024 (2013.01); C12N 15/1037 (2013.01); C12N 15/1068 (2013.01); C12N 15/1093 (2013.01); C12N 15/907 (2013.01); C07K 2317/14 (2013.01); C07K 2317/51 (2013.01); C07K 2317/515 (2013.01); C07K 2317/622 (2013.01); C40B 30/04 (2013.01); C40B 50/06 (2013.01);
Abstract

Disclosed herein is an efficient method of generating a library of variants of a sequence of interest, such as may be used in directed evolution, in one embodiment, the method includes an amplification reaction, e.g. error-prone PCR, to generate double-stranded DNA (dsDNA) variants of a sequence of interest, after which one strand of the dsDNA variants may be selectively degraded to produce single-stranded DNA (ssDNA) variants. The ssDNA variants may be hybridized to ssDNA intermediaries, e.g., uracilated circular ssDNA intermediaries, to form heteroduplex DNA, which may be transformed into cells, such ascells, yielding a library of variants. This method eliminates the inefficient sub-cloning steps and the need for costly primer sets required by many prior methods.


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