US 10,138,512 B2
Nucleic acid processing of a nucleic acid fragment with a triazole linkage
Afaf H. El-Sagheer, Hampshire (GB); and Tom Brown, Hampshire (GB)
Assigned to ATDBIO LIMITED, Southhampton (GB)
Appl. No. 15/312,313
Filed by ATDBio Limited, Southampton (GB)
PCT Filed May 18, 2015, PCT No. PCT/GB2015/051451
§ 371(c)(1), (2) Date Nov. 18, 2016,
PCT Pub. No. WO2015/177520, PCT Pub. Date Nov. 26, 2015.
Claims priority of application No. 1408841.3 (GB), filed on May 19, 2014.
Prior Publication US 2017/0088888 A1, Mar. 30, 2017
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6855 (2018.01); C12Q 1/6853 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/6869 (2018.01)
CPC C12Q 1/6855 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6853 (2013.01); C12Q 1/6869 (2013.01)] 19 Claims
1. A method of nucleic acid processing comprising:
providing an adapted nucleic acid fragment comprising a nucleic acid fragment linked at its 3′-end to a 3′-adapter molecule by a triazole linkage comprising a 1,2,3-triazole group, wherein the 3′-adapter molecule comprises RNA, DNA, a nucleic acid analogue or combinations thereof;
annealing a primer to the adapted nucleic acid fragment, wherein the primer anneals to a region of the 3′-adapter molecule; and
processing the adapted nucleic acid fragment by transcribing at least a portion of the adapted nucleic acid fragment with reverse transcriptase, wherein said portion of the adapted nucleic acid fragment transcribed with reverse transcriptase comprises said triazole linkage.