US 11,807,956 B2
Oligonucleic acid variant libraries and synthesis thereof
Anthony Cox, Mountain View, CA (US); Sebastian Treusch, San Francisco, CA (US); and Siyuan Chen, San Mateo, CA (US)
Assigned to Twist Bioscience Corporation, South San Francisco, CA (US)
Filed by Twist Bioscience Corporation, South San Francisco, CA (US)
Filed on Oct. 12, 2020, as Appl. No. 17/068,551.
Application 17/068,551 is a continuation of application No. 15/268,422, filed on Sep. 16, 2016, granted, now 10,844,373.
Claims priority of provisional application 62/354,034, filed on Jun. 23, 2016.
Claims priority of provisional application 62/263,548, filed on Dec. 4, 2015.
Claims priority of provisional application 62/220,879, filed on Sep. 18, 2015.
Prior Publication US 2021/0040476 A1, Feb. 11, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C40B 50/06 (2006.01); C12N 15/10 (2006.01); G01N 33/50 (2006.01); C40B 50/14 (2006.01); C40B 50/08 (2006.01); C12N 15/113 (2010.01); C12Q 1/6876 (2018.01); C12Q 1/6886 (2018.01)
CPC C40B 50/06 (2013.01) [C12N 15/1079 (2013.01); C12N 15/1093 (2013.01); C12N 15/113 (2013.01); C12Q 1/6876 (2013.01); C40B 50/08 (2013.01); C40B 50/14 (2013.01); G01N 33/502 (2013.01); G01N 33/5029 (2013.01); G01N 33/5041 (2013.01); C12N 2320/30 (2013.01); C12N 2330/31 (2013.01); C12Q 1/6886 (2013.01); C12Q 2600/158 (2013.01)] 16 Claims
OG exemplary drawing
 
1. A method for generating a library of variant nucleic acids, the method comprising:
a) providing predetermined nucleic acid sequences, encoding for at least about 30,000 non-identical oligonucleotides, wherein each of the at least about 30,000 non-identical oligonucleotides encodes for a variant codon sequence compared to a single reference sequence, wherein the predetermined nucleic acid sequences encode for 19 variants for each position of multiple codon positions compared to the single reference sequence;
b) providing a structure having a surface;
c) synthesizing the at least about 30,000 non-identical oligonucleotides, wherein each of the at least about 30,000 non-identical oligonucleotides extends from the surface; and
d) mixing the at least about 30,000 non-identical oligonucleotides with a DNA polymerase and the single reference sequence to form a library of variant nucleic acids by amplification.