公开(公告)号：US10407720B2

公开(公告)日：2019-09-10

申请号：US15037322

申请日：2014-12-14

Applicant: ACADEMIA SINICA

Inventor： Kuo Ping Chiu ,  Yu-Shin Nai

IPC: C12Q1/686 ,  G16B30/00 ,  C12Q1/6869 ,  C12Q1/6886 ,  C12Q1/6855 ,  G16B35/00 ,  G16C20/60

Abstract: An adaptor for use in amplifying all linear, double-stranded nucleic acid molecules of unknown sequences in a sample is disclosed. The adaptor consists of: (1) the first oligonucleotide (P-oligo) with a phosphate at the 5′ end and without an additional thymine nucleotide at the 3′ end; and (2) the second oligonucleotide (T˜oligo) with an extra 3′-T and without a 5′-phosphate. The P-oligo and T-oligo are complementary to each other except at the 3′-T (thymine) in the T-oligo. The adaptor is ligated to nucleic acids of unknown sequences which have an extra A in the 3′ end (3′˜A overhang) to form adaptor-ligated target nucleic acids. The T-oligo is then employed as a single primer for T-oligo-primed polymerase chain reaction (TOP-PCR) and amplifies the nucleic acids of unknown sequences in full-length.