公开(公告)号：US20160298172A1

公开(公告)日：2016-10-13

申请号：US15037322

申请日：2014-12-14

Applicant: ACADEMIA SINICA

Inventor： Kuo Ping CHIU ,  Yu-Shin NAI

IPC: C12Q1/68 ,  G06F19/22 ,  C40B30/02

CPC classification number: C12Q1/686 ,  C12Q1/6855 ,  C12Q1/6869 ,  C12Q1/6886 ,  C12Q2600/156 ,  G16B30/00 ,  G16B35/00 ,  G16C20/60 ,  C12Q2521/501 ,  C12Q2525/155 ,  C12Q2525/173 ,  C12Q2525/191 ,  C12Q2531/113 ,  C12Q2535/122

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.

Abstract translation: 公开了用于扩增样品中未知序列的所有线性双链核酸分子的衔接子。 衔接头由以下部分组成：（1）在5'末端具有磷酸酯的第一寡核苷酸（P-oligo），在3'端没有另外的胸腺嘧啶核苷酸; 和（2）具有额外的3'-T并且没有5'-磷酸的第二寡核苷酸（T〜ol）。 除了在T-oligo中的3'-T（胸腺嘧啶）之外，P-寡聚和T-寡聚是互补的。 将衔接子连接到在3'末端具有额外A（3'-A突出端）的未知序列的核酸以形成衔接物连接的靶核酸。 然后将T-oligo用作T-oligo-primed聚合酶链反应（TOP-PCR）的单引物，并扩增全长未知序列的核酸。