	US 11,697,843 B2
	Methods for creating directional bisulfite-converted nucleic acid libraries for next generation sequencing
Benjamin G. Schroeder, San Mateo, CA (US); and Doug Amorese, Los Altos, CA (US)
Assigned to Tecan Genomics, Inc., Redwood City, CA (US)
Filed by Tecan Genomics, Inc., Redwood City, CA (US)
Filed on May 14, 2021, as Appl. No. 17/321,030.
Application 17/321,030 is a continuation of application No. 14/991,340, filed on Jan. 8, 2016, granted, now 11,028,430.
Application 14/991,340 is a continuation of application No. 13/938,059, filed on Jul. 9, 2013, abandoned.
Claims priority of provisional application 61/801,382, filed on Mar. 15, 2013.
Claims priority of provisional application 61/669,613, filed on Jul. 9, 2012.
Prior Publication US 2021/0285040 A1, Sep. 16, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6858 (2018.01); C12Q 1/6855 (2018.01)

CPC C12Q 1/6858 (2013.01) [C12Q 1/6855 (2013.01)]

20 Claims

1. A method for generating a nucleic acid library, the method comprising:

ligating first strands of oligonucleotide adapters of similar construct to 5′ ends of double-stranded DNA fragments in a sample, wherein the first strand of each adapter comprises one or more guanine;

extending 3′ ends of the double-stranded DNA fragments with a DNA polymerase in the presence of a dCTP analog resistant to bisulfite treatment, thereby creating extension products comprising one or more cytosines resistant to bisulfite treatment and complementary to the guanines;

treating the extension products with bisulfite, thereby converting unmodified cytosines bases in the DNA fragments and adapter sequences to uracil; and

amplifying the bisulfite-treated extension products.