公开(公告)号：US09920360B2

公开(公告)日：2018-03-20

申请号：US14233711

申请日：2013-11-04

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Linda Wong ,  Caifu Chen ,  Yalei Wu ,  Shoulian Dong ,  Chunmei Liu

IPC: C12Q1/68 ,  C12P19/34

CPC classification number: C12Q1/6855 ,  C12P19/34 ,  C12Q1/6832 ,  C12Q1/6848 ,  C12Q1/6851 ,  C12Q1/6853 ,  C12Q1/6858 ,  C12Q1/686 ,  C12Q2525/191 ,  C12Q2525/301

Abstract: Methods, compositions and kits for capturing, detecting and quantifying mature small RNAs are provided herein. Embodiments of the methods comprise ligating 5′ and 3′ ligation adaptors to the 5′ and 3′ ends of the mature small RNAs, respectively, in the presence of 5′ and 3′ semi-degenerate ligation splints to generate a ligation product. Other embodiments comprise reverse transcribing polyadenylated mature small RNA with a universal reverse transcription primer and ligating an adaptor to the 3′ end of the cDNA in the presence of a semi-degenerate ligation splint to generate a cDNA ligation product.

公开(公告)号：US11208688B2

公开(公告)日：2021-12-28

申请号：US15887159

申请日：2018-02-02

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Linda Wong ,  Caifu Chen ,  Yalei Wu ,  Shoulian Dong ,  Chunmei Liu

IPC: C12Q1/6855 ,  C12P19/34 ,  C12Q1/6851 ,  C12Q1/6858 ,  C12Q1/686 ,  C12Q1/6853 ,  C12Q1/6832 ,  C12Q1/6848

Abstract: Methods, compositions and kits for capturing, detecting and quantifying mature small RNAs are provided herein. Embodiments of the methods comprise ligating 5′ and 3′ ligation adaptors to the 5′ and 3′ ends of the mature small RNAs, respectively, in the presence of 5′ and 3′ semi-degenerate ligation splints to generate a ligation product. Other embodiments comprise reverse transcribing polyadenylated mature small RNA with a universal reverse transcription primer and ligating an adaptor to the 3′ end of the cDNA in the presence of a semi-degenerate ligation splint to generate a cDNA ligation product.

公开(公告)号：US11274340B2

公开(公告)日：2022-03-15

申请号：US16741471

申请日：2020-01-13

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Chunmei Liu ,  Shoulian Dong ,  Linda Wong

IPC: C12Q1/686 ,  C12Q1/6806 ,  C12Q1/6851

Abstract: Methods, compositions and kits for capturing, detecting and quantifying mature small RNAs are provided herein. Embodiments of the methods comprise tailing both the 5′ and 3′ ends of mature small RNA by ligating a 5′ ligation adaptor to the 5′ end and polyadenylating the 3′ end. Other embodiments comprise reverse transcribing the adaptor ligated, polyadenylated mature small RNA with a universal reverse transcription primer and amplifying the cDNA with universal primers.

公开(公告)号：US10563250B2

公开(公告)日：2020-02-18

申请号：US15066139

申请日：2016-03-10

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Chunmei Liu ,  Shoulian Dong ,  Linda Wong

IPC: C12P19/34 ,  C12Q1/686

Abstract: Methods, compositions and kits for capturing, detecting and quantifying mature small RNAs are provided herein. Embodiments of the methods comprise tailing both the 5′ and 3′ ends of mature small RNA by ligating a 5′ ligation adaptor to the 5′ end and polyadenylating the 3′ end. Other embodiments comprise reverse transcribing the adaptor ligated, polyadenylated mature small RNA with a universal reverse transcription primer and amplifying the cDNA with universal primers.

公开(公告)号：US20150105275A1

公开(公告)日：2015-04-16

申请号：US14233711

申请日：2013-11-04

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Linda Wong ,  Caifu Chen ,  Yalei Wu ,  Shoulian Dong ,  Chunmei Liu

IPC: C12Q1/68

CPC classification number: C12Q1/6855 ,  C12P19/34 ,  C12Q1/6832 ,  C12Q1/6848 ,  C12Q1/6851 ,  C12Q1/6853 ,  C12Q1/6858 ,  C12Q1/686 ,  C12Q2525/191 ,  C12Q2525/301

Abstract: Methods, compositions and kits for capturing, detecting and quantifying mature small RNAs are provided herein. Embodiments of the methods comprise ligating 5′ and 3′ ligation adaptors to the 5′ and 3′ ends of the mature small RNAs, respectively, in the presence of 5′ and 3′ semi-degenerate ligation splints to generate a ligation product. Other embodiments comprise reverse transcribing polyadenylated mature small RNA with a universal reverse transcription primer and ligating an adaptor to the 3′ end of the cDNA in the presence of a semi-degenerate ligation splint to generate a cDNA ligation product.

Abstract translation: 本文提供了用于捕获，检测和定量成熟小RNA的方法，组合物和试剂盒。 方法的实施方案包括在5'和3'半简并连接夹板的存在下分别将5'和3'连接衔接子连接到成熟小RNA的5'和3'末端以产生连接产物。 其他实施方案包括具有通用逆转录引物的逆转录聚腺苷酸化成熟小RNA，并且在半退化连接夹板存在下将衔接子连接到cDNA的3'末端以产生cDNA连接产物。