公开(公告)号：US10472671B2

公开(公告)日：2019-11-12

申请号：US14955386

申请日：2015-12-01

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Shiaw-Min Chen ,  Elana Swartzman ,  David Ruff ,  Mark Shannon ,  Julia Lu ,  Stephen Hendricks

IPC: C12Q1/68 ,  C12Q1/6855 ,  C12Q1/6804

Abstract: This application relates to methods for ligating oligonucleotides having complementarity to a target nucleic acid, and amplifying the ligated oligonucleotides, where ligation and amplification occur in the same reaction mixture.

公开(公告)号：US09334531B2

公开(公告)日：2016-05-10

申请号：US14023361

申请日：2013-09-10

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Chieh-Yuan Li ,  David Ruff ,  Shiaw-Min Chen ,  Jennifer O'Neil ,  Rachel Kasinskas ,  Jonathan Rothberg ,  Bin Li ,  Kai Qin Lao

IPC: C12Q1/68

CPC classification number: C12N9/1252 ,  C12Q1/6846 ,  C12Q1/6853 ,  C12Q1/6855 ,  C12Q1/686 ,  C12Q1/6874 ,  C12Y207/07007 ,  C12Q2531/119 ,  C12Q2565/537

Abstract: In some embodiments, the present teachings provide methods for nucleic acid amplification, comprising forming a reaction mixture, and subjecting the reaction mixture to conditions suitable for nucleic acid amplification. In some embodiments, methods for nucleic acid amplification include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, methods for nucleic acid amplification include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the methods for nucleic acid amplification employ an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel. In some embodiments, methods for nucleic acid amplification can be conducted in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components. In some embodiments, methods for nucleic acid amplification comprise a amplifying at least one polynucleotide onto a surface under isothermal amplification conditions, optionally in the presence of a polymer. The polymer can include a sieving agent and/or a diffusion-reducing agent.