公开(公告)号：EP3095879B1

公开(公告)日：2018-09-19

申请号：EP16177421.1

申请日：2013-03-15

Applicant: Life Technologies Corporation

Inventor： LI, Chieh-Yuan ,  RUFF, David ,  CHEN, Shiaw-Min ,  O'NEIL, Jennifer ,  KASINSKAS, Rachel ,  ROTHBERG, Jonathan

IPC: C12Q1/68

CPC classification number: C12Q1/6806 ,  C12Q2521/507 ,  C12Q2527/101 ,  C12Q2527/153 ,  C12Q2535/122 ,  C12Q2563/155 ,  C12Q2563/159 ,  C12Q2565/537 ,  C12Q2565/607

Abstract: In some embodiments, the present teachings provide methods for nucleic acid amplification, comprising forming an 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 an 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.

公开(公告)号：EP2539473A2

公开(公告)日：2013-01-02

申请号：EP11748228.1

申请日：2011-02-28

Applicant: Life Technologies Corporation

Inventor： DAVIDSON, John, F. ,  HINZ, Wolfgang ,  ROTHBERG, Jonathan ,  WHITAKER, Richard

IPC: C12Q1/68 ,  C12Q1/48 ,  C12N9/12

CPC classification number: C12N9/1252

Abstract: Methods, compositions, systems, apparatuses and kits comprising modified proteins, particularly modified nucleic acid-binding proteins with altered buffering properties are provided. For example, in some embodiments, methods of forming modified proteins including one or more amino acid modifications to achieve desired pKa values are described. Furthermore, the invention provides methods for using such modified proteins in ion-producing reactions, such as ion-based nucleic acid sequencing reactions.

Abstract translation: 提供了包含修饰的蛋白质，特别是具有改变的缓冲性质的修饰的核酸结合蛋白的方法，组合物，系统，装置和试剂盒。 例如，在一些实施方案中，描述了形成包含一个或多个氨基酸修饰的修饰蛋白以实现期望的pKa值的方法。 此外，本发明提供了在离子生成反应中使用这些修饰蛋白的方法，如基于离子的核酸测序反应。