US 11,739,367 B2
Reagents and adapters for nucleic acid sequencing and methods for making such reagents and adapters
Jesse J. Salk, Seattle, WA (US); and Scott R. Kennedy, Seattle, WA (US)
Assigned to TwinStrand Biosciences, Inc., Seattle, WA (US)
Appl. No. 16/758,726
Filed by TwinStrand Biosciences, Inc., Seattle, WA (US)
PCT Filed Nov. 8, 2018, PCT No. PCT/US2018/059908
§ 371(c)(1), (2) Date Apr. 23, 2020,
PCT Pub. No. WO2019/094651, PCT Pub. Date May 16, 2019.
Claims priority of provisional application 62/583,462, filed on Nov. 8, 2017.
Prior Publication US 2020/0362390 A1, Nov. 19, 2020
Int. Cl. C12Q 1/6806 (2018.01); C12Q 1/6869 (2018.01)
CPC C12Q 1/6806 (2013.01) [C12Q 1/6869 (2013.01)] 15 Claims
 
1. A method for preparing a duplex adapter, comprising:
annealing an elongation strand and a template strand at a complementary region to form a first intermediate duplex adapter product, wherein the template strand comprises:
an identifier sequence,
a first capture label attached to the template strand via a first cut site template, and
a second capture label attached to the template strand via a second cut site;
extending the elongation strand to at least partially duplicate the identifier sequence to form a second intermediate duplex adapter product comprising a first cut site between the identifier sequence and the first capture label;
cutting the second intermediate duplex adapter product at the first cut site to form a third intermediate duplex adapter product and a first cleaved by-product comprising the first capture label;
removing undesired products comprising the first capture label; and
cutting at the second cut site to form the duplex adapter and a second cleaved by-product comprising the second capture label.