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:
Oct. 25, 2016

Filed:

Jul. 31, 2008
Applicant:

Xuqiu Tan, San Diego, CA (US);

Inventor:

Xuqiu Tan, San Diego, CA (US);

Assignee:

BASF Enzymes LLC, San Diego, CA (US);

Attorneys:
Primary Examiner:
Int. Cl.
CPC ...
C12P 19/34 (2006.01); C12N 15/10 (2006.01);
U.S. Cl.
CPC ...
C12P 19/34 (2013.01); C12N 15/102 (2013.01); C12N 15/1024 (2013.01);
Abstract

The present invention provides a novel method of producing a plurality of modified polynucleotides having different combinations of various mutations at multiple sites by a tailored multi-site combinatorial assembly, comprising adding at least two or at least three primers to a double stranded template polynucleotide in a single reaction mixture, wherein the primers are not overlapping, and wherein each of the primers comprise at least one mutation different from the other primers, wherein at least one primer is a forward primer that can anneal to a minus strand of the template and at least one primer is a reverse primer that can anneal to a plus strand of the template, and subjecting the reaction mixture to a polymerase extension reaction to yield a plurality of extended modified polynucleotides from the at least three primers. The method can be performed without employing a ligation step prior to transforming the extended modified polynucleotides into a cell. The plurality of extended modified polynucleotides can be treated with an enzyme for destroying the template polynucleotide prior to transforming in to the cell.


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