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：US11254977B2

公开(公告)日：2022-02-22

申请号：US14206995

申请日：2014-03-12

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Kelly Li ,  Shoulian Dong ,  Caifu Chen

IPC: C12Q1/68 ,  C12Q1/6858

Abstract: The present disclosure is drawn to methods for detection, quantitation and analysis of nucleotides of interest, for example SNPs, in nucleic acid sequences of interest using universal FRET-based reporter primers.

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：US10689694B2

公开(公告)日：2020-06-23

申请号：US16109191

申请日：2018-08-22

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Caifu Chen ,  Ruoying Tan

IPC: C12Q1/68 ,  C12Q1/6858 ,  C12Q1/6883 ,  C12Q1/686

Abstract: In some embodiments, the present inventions relates generally to compositions, methods and kits for use in discriminating sequence variation between different alleles. More specifically, in some embodiments, the present invention provides for compositions, methods and kits for quantitating rare (e.g., mutant) allelic variants, such as SNPs, or nucleotide (NT) insertions or deletions, in samples comprising abundant (e.g., wild type) allelic variants with high specificity and selectivity. In particular, in some embodiments, the invention relates to a highly selective method for mutation detection referred to as competitive allele-specific TaqMan PCR (“cast-PCR”).

公开(公告)号：US09512473B2

公开(公告)日：2016-12-06

申请号：US14445046

申请日：2014-07-28

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Caifu Chen ,  Ruoying Tan

IPC: C12Q1/68

CPC classification number: C12Q1/6858 ,  C12Q1/686 ,  C12Q1/6883 ,  C12Q2600/156 ,  C12Q2561/113 ,  C12Q2561/101 ,  C12Q2537/161 ,  C12Q2525/186 ,  C12Q2565/107 ,  C12Q2535/125

Abstract: In some embodiments, the present inventions relates generally to compositions, methods and kits for use in discriminating sequence variation between different alleles. More specifically, in some embodiments, the present invention provides for compositions, methods and kits for quantitating rare (e.g., mutant) allelic variants, such as SNPs, or nucleotide (NT) insertions or deletions, in samples comprising abundant (e.g., wild type) allelic variants with high specificity and selectivity. In particular, in some embodiments, the invention relates to a highly selective method for mutation detection referred to as competitive allele-specific TaqMan PCR (“cast-PCR”).

Abstract translation: 在一些实施方案中，本发明一般涉及用于区分不同等位基因之间的序列变异的组合物，方法和试剂盒。 更具体地说，在一些实施方案中，本发明提供了用于定量稀少（例如，突变体）等位基因变体如SNP或核苷酸（NT）插入或缺失的组合物，方法和试剂盒，其包含丰富的（例如，野生型 ）等位基因变体，具有高特异性和选择性。 特别地，在一些实施方案中，本发明涉及用于突变检测的高选择性方法，称为竞争性等位基因特异性TaqMan PCR（“cast-PCR”）。

公开(公告)号：US20150099644A1

公开(公告)日：2015-04-09

申请号：US14395456

申请日：2013-04-19

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Caifu Chen ,  Casey McFarland ,  David Keys

IPC: C12Q1/68

CPC classification number: C12Q1/686 ,  C12Q1/6851 ,  C12Q1/6858 ,  C12Q2527/119 ,  C12Q2563/159 ,  C12Q2565/607 ,  C12Q2565/301

Abstract: A method of detection of a target nucleic acid is provided. The method includes fractionating a sample into a plurality of sample volumes wherein more than 50% of the fractions contain no more than 1 target nucleic acid molecule per sample volumes, and subjecting the plurality of sample volumes to conditions for amplification. The method further includes detecting a change in ion concentration in a sample volume wherein a target nucleic acid is present, counting the number of fractions with an amplified target nucleic acid, and determining the quantity of target nucleic acid in the sample.

Abstract translation: 提供了检测靶核酸的方法。 该方法包括将样品分馏成多个样品体积，其中超过50％的级分每个样品体积含有不超过1个靶核酸分子，并且对多个样品体积进行扩增条件。 该方法还包括检测样品体积中的离子浓度的变化，其中存在靶核酸，用扩增的靶核酸计数分数，并测定样品中靶核酸的量。