US 12,071,660 B2
Bisulfite-free, base-resolution identification of cytosine modifications
Chunxiao Song, Oxford (GB); and Yibin Liu, Oxford (GB)
Assigned to Ludwig Institute for Cancer Research LTD., Zurich (CH)
Appl. No. 16/960,510
Filed by Ludwig Institute for Cancer Research Ltd, Zurich (CH)
PCT Filed Jan. 8, 2019, PCT No. PCT/US2019/012627
§ 371(c)(1), (2) Date Jul. 7, 2020,
PCT Pub. No. WO2019/136413, PCT Pub. Date Jul. 11, 2019.
Claims priority of provisional application 62/771,409, filed on Nov. 26, 2018.
Claims priority of provisional application 62/660,523, filed on Apr. 20, 2018.
Claims priority of provisional application 62/614,798, filed on Jan. 8, 2018.
Prior Publication US 2020/0370114 A1, Nov. 26, 2020
Int. Cl. C12Q 1/68 (2018.01); C12P 17/16 (2006.01); C12Q 1/6844 (2018.01); C12Q 1/6869 (2018.01); C12Q 1/6874 (2018.01); C12Q 1/6876 (2018.01)
CPC C12Q 1/6869 (2013.01) [C12P 17/16 (2013.01); C12Q 1/6844 (2013.01); C12Q 1/6874 (2013.01); C12Q 1/6876 (2013.01); C12Q 2600/154 (2013.01); C12Y 204/01026 (2013.01); C12Y 204/01027 (2013.01)] 19 Claims
 
1. A method comprising steps of:
Providing a nucleic acid sample comprising one or more 5-carboxylcytosine (5caC) or 5-formylcytosine (5fC) residues; and
Contacting the sample with a borane reducing agent under conditions that reduce 5caC and 5fC to dihydrouracil (DHU),
wherein the nucleic acid sample comprising one or more 5caC or 5fC residues comprises ligated nucleic acids.