公开(公告)号：US20190119737A1

公开(公告)日：2019-04-25

申请号：US16301687

申请日：2017-05-16

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Janay Kong ,  Aydogan Ozcan ,  Omai Garner

IPC: C12Q1/6844 ,  C09B29/16 ,  C09B23/04 ,  C09B15/00 ,  C09B69/04

Abstract: A fluorescent dye and quencher mixture for reporting on nucleic acid amplification from a sample includes a fluorescent intercalating dye, a dye sequestering or quenching agent such as hydroxynapthol blue (HNB) or caffeine, and primers, dNTPs, and a nucleic acid polymerizing enzyme or fragment thereof. The presence of the dye in combination with the dye sequestering or quenching agent improves the overall dynamic range of the fluorescent signal as well as shortens the time needed for visualization or image capture of amplified nucleic acid. The fluorescent dye and quencher mixture also enables the detection of nucleic acids in samples having low copy numbers.

公开(公告)号：US20160257990A1

公开(公告)日：2016-09-08

申请号：US15029231

申请日：2014-10-17

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Janay Kong

IPC: C12Q1/68

CPC classification number: C12Q1/6804 ,  C12Q1/6844 ,  C12Q2563/159 ,  C12Q2563/173

Abstract: A method of detecting a target and quantifying the concentration of the same within a sample includes generating a plurality of fractionated volumes, wherein at least some of the fractionated volumes contain the target bound to a first detector molecule connected to a first reaction component (R1) and a second detector molecule connected to a second reaction component (R2). The fractionated volumes that contain the target, first detector molecule, second detector molecule, and a probe or other reporter molecule emit light and are imaged. Fractionated volumes emitting radiation can be used to detect the presence of the target within the sample. The number of fractionated volumes emitting a positive emission signal can be counted from the image and the concentration (or range of calculations) of the target can be calculated based at least in part on the number of fractionated volumes emitting a positive emission signal from the image.

Abstract translation: 检测靶标并在样品中定量其浓度的方法包括产生多个分馏体积，其中至少部分分馏体积包含与连接到第一反应组分（R1）的第一检测分子结合的靶标， 和连接到第二反应组分（R2）的第二检测器分子。 包含目标，第一检测器分子，第二检测器分子和探针或其他报告分子的分级体积发光并成像。 可以使用发射辐射的分级体积来检测样品中目标物的存在。 可以从图像中计数发射正发射信号的分级体积的数量，并且可以至少部分地基于从图像发射正发射信号的分级体积的数量来计算目标的浓度（或计算范围） 。