公开(公告)号：US20200248166A1

公开(公告)日：2020-08-06

申请号：US16837674

申请日：2020-04-01

申请人： RGENE, Inc.

发明人： Yu Zheng ,  Manqing Hong

IPC分类号： C12N9/00

摘要： Provided herein are compositions, systems, and methods using multiple ligases, wherein at least one of the ligases is an adenylation-deficient ATP-dependent ligase or an un-adenylated ATP-dependent ligase (e.g., present in an ATP free mixture). In certain embodiments, multiple ligases are used to ligate a pre-adenylated double stranded sequence to a non-adenylated double stranded sequence (e.g., the adenylation-deficient ATP-dependent ligase or un-adenylated ATP-dependent ligase ligates the first strand, and a second ligase ligates the second strand). In other embodiments, provided herein are mutant T4 ligases (e.g., K159S mutant or K159C mutant).

公开(公告)号：US11124789B2

公开(公告)日：2021-09-21

申请号：US16837674

申请日：2020-04-01

申请人： RGENE, Inc.

发明人： Yu Zheng ,  Manqing Hong

IPC分类号： C12N9/00

摘要： Provided herein are compositions, systems, and methods using multiple ligases, wherein at least one of the ligases is an adenylation-deficient ATP-dependent ligase or an un-adenylated ATP-dependent ligase (e.g., present in an ATP free mixture). In certain embodiments, multiple ligases are used to ligate a pre-adenylated double stranded sequence to a non-adenylated double stranded sequence (e.g., the adenylation-deficient ATP-dependent ligase or un-adenylated ATP-dependent ligase ligates the first strand, and a second ligase ligates the second strand). In other embodiments, provided herein are mutant T4 ligases (e.g., K159S mutant or K159C mutant).

公开(公告)号：US20190062827A1

公开(公告)日：2019-02-28

申请号：US16082500

申请日：2017-03-14

申请人： RGENE, Inc.

发明人： Yu Zheng ,  Manqing Hong

IPC分类号： C12Q1/6869 ,  C12N9/00 ,  C12Q1/686 ,  C07H21/04

摘要： Provided herein are compositions, systems, and methods employing hyper-thermostable lysine-mutant ssDNA/RNA ligases that possesses both ssRNA ligase and ssDNA ligase activity. In certain embodiments, such hyper-thermostable lysine-mutant ssDNA/RNA ligases are used to ligate an first single stranded nucleic acid sequence with a 5′ adenylated end to a second single stranded nucleic acid sequence (e.g., at a temperature of at least 75° C.) to form a ligated nucleic acid sequence. In further embodiments, the ligated nucleic acid sequence is sequenced.

公开(公告)号：US20170226498A1

公开(公告)日：2017-08-10

申请号：US15426543

申请日：2017-02-07

申请人： RGENE, Inc.

发明人： Yu Zheng ,  Manqing Hong

IPC分类号： C12N9/00

CPC分类号： C12N9/93 ,  C12Y605/01001 ,  C12Y605/01002

摘要： Provided herein are compositions, systems, and methods using multiple ligases, wherein at least one of the ligases is an adenylation-deficient ATP-dependent ligase or an un-adenylated ATP-dependent ligase (e.g., present in an ATP free mixture). In certain embodiments, multiple ligases are used to ligate a pre-adenylated double stranded sequence to a non-adenylated double stranded sequence (e.g., the adenylation-deficient ATP-dependent ligase or un-adenylated ATP-dependent ligase ligates the first strand, and a second ligase ligates the second strand). In other embodiments, provided herein are mutant T4 ligases (e.g., K159S mutant or K159C mutant).