Patent ID: 11920129
Assignee: NATIONAL UNIVERSITY OF SINGAPORE
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 3:
4. The method according to claim 1; wherein step (i) comprises:
(i)(a) providing a double-stranded nucleic acid template comprising a first nucleic acid strand and a second nucleic acid strand substantially reverse complementary to the first nucleic acid strand;
(i)(b) providing a first primer comprising a first sequence with at least one modified nucleotide upstream of a second sequence substantially complementary to the second strand of the nucleic acid template; and a second primer comprising a third sequence with at least one modified nucleotide upstream of a fourth sequence substantially complementary to the first strand of the nucleic acid template;
(i)(c) amplifying the nucleic acid template using the first and second primers to produce an amplicon; and
(i)(d) chemically cleaving the amplicon to produce the first nucleic acid molecule comprising a first overhang of one nucleotide in length at a first end and a second overhang of at least one nucleotide of one nucleotide in length at its other (or second) end; wherein the first overhang and the second overhang are different nucleotides and are not complementary to each other; or
(i)(a) providing a first single-stranded nucleic acid molecule;
(i)(b) providing a second single-stranded nucleic acid molecule substantially complementary to the first single nucleic acid molecule; and
(i)(c) allowing the first and second single-stranded nucleic acid molecule to anneal to produce the first nucleic acid molecule comprising a first overhang of one nucleotide in length at a first end and a second overhang of one nucleotide in length at its other (or second) end; wherein the first overhang and the second overhang are different nucleotides and are not complementary to each other.