公开(公告)号：WO2014071322A1

公开(公告)日：2014-05-08

申请号：PCT/US2013/068350

申请日：2013-11-04

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： WONG, Linda ,  CHEN, Caifu ,  WU, Yalei ,  DONG, Shoulian ,  LIU, Chunmei

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 lligation 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多聚腺苷酸成熟小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。

公开(公告)号：WO2015153973A1

公开(公告)日：2015-10-08

申请号：PCT/US2015/024249

申请日：2015-04-03

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： WU, Yalei ,  KEYS, David

IPC: C12Q1/68

CPC classification number: G06F19/24 ,  C12Q1/6851 ,  G06F19/12 ,  G06F19/20 ,  G06F19/22 ,  G06F19/26 ,  C12Q2537/16 ,  C12Q2545/114

Abstract: A method for quantifying nucleic acid is provided. The method includes determining a first reference threshold cycle for a first predetermined input quantity for a reference nucleic acid, determining a first target threshold cycle for the first predetermined input quantity for a target nucleic acid, determining a second reference threshold cycle for a second predetermined input quantity for the reference nucleic acid, and determining a second target threshold cycle, by the processor, for the second predetermined input quantity for the target nucleic acid. The method further includes receiving a sample threshold cycle, determining a sample input quantity based on the first and second reference threshold cycle and the first and second target threshold cycle, and displaying the sample input quantity to a user.

Abstract translation: 提供了定量核酸的方法。 该方法包括确定用于参考核酸的第一预定输入量的第一参考阈值循环，确定靶核酸的第一预定输入量的第一目标阈值循环，为第二预定输入确定第二参考阈值循环 参考核酸的量，以及由处理器确定靶核酸的第二预定输入量的第二目标阈值循环。 该方法还包括接收采样阈值周期，基于第一和第二参考阈值周期以及第一和第二目标阈值周期来确定采样输入量，以及将样本输入量显示给用户。