US 12,247,252 B2
Method of improved sequencing by strand identification
Malek Faham, Burlingame, CA (US); Shengrong Lin, Fremont, CA (US); Ling Fung Tang, San Francisco, CA (US); Yontao Lu, Sunnyvale, CA (US); Zhaohui Sun, Albany, CA (US); Yingyu Wang, Mountain View, CA (US); and Li Weng, Fremont, CA (US)
Assigned to ACCURAGEN HOLDINGS LIMITED, George Town (KY)
Filed by ACCURAGEN HOLDINGS LIMITED, George Town (KY)
Filed on Jul. 28, 2022, as Appl. No. 17/815,815.
Application 17/815,815 is a continuation of application No. 16/301,707, granted, now 11,427,866, previously published as PCT/US2017/032980, filed on May 16, 2017.
Claims priority of provisional application 62/506,390, filed on May 15, 2017.
Claims priority of provisional application 62/337,296, filed on May 16, 2016.
Prior Publication US 2023/0167493 A1, Jun. 1, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6869 (2018.01); C12Q 1/6827 (2018.01); C12Q 1/6858 (2018.01); C40B 50/06 (2006.01)
CPC C12Q 1/6869 (2013.01) [C12Q 1/6827 (2013.01); C12Q 1/6858 (2013.01); C40B 50/06 (2013.01); C12Q 2521/501 (2013.01); C12Q 2535/139 (2013.01); C12Q 2537/143 (2013.01)] 8 Claims
 
1. A method of identifying complementary strands in a nucleic acid sample comprising a plurality of double-stranded polynucleotides, each double-stranded polynucleotide of the plurality comprising a first complementary strand and a second complementary strand, each having a 5′ end and a 3′ end, the method comprising:
(a) modifying a polynucleotide sequence of at least one of a first complementary strand and a second complementary strand of individual double-stranded polynucleotides by adding one or more predetermined nucleotides in a base by base manner to the 3′ end of at least one of the first complementary strand and the second complementary strand to yield a plurality of modified double-stranded polynucleotides;
(b) sequencing said plurality of modified double-stranded polynucleotides or a derivative thereof to yield a plurality of sequencing reads; and
(c) identifying from said plurality of sequencing reads, a given first complementary strand and a given second complementary strand as originating from a common double-stranded polynucleotide based on (i) sequences of the 5′ ends, (ii) sequences 3′ ends adjacent to the one or more predetermined nucleotides, and (iii) sequences of the one or more predetermined nucleotides.