公开(公告)号：US09593365B2

公开(公告)日：2017-03-14

申请号：US14434274

申请日：2013-10-16

Applicant: SPATIAL TRANSCRIPTOMICS AB

Inventor： Jonas Frisen ,  Patrik Stahl ,  Joakim Lundeberg ,  Fredrik Salmen

IPC: C12Q1/68

CPC classification number: C12Q1/6837 ,  C12Q1/6841 ,  C12Q1/6874 ,  C12Q2543/101 ,  C12Q2565/514 ,  C12Q2565/537

Abstract: The present invention relates to methods and products for localized or spatial detection and/or analysis of RNA in a tissue sample or a portion thereof, comprising: (a) providing an object substrate on which at least one species of capture probe, comprising a capture domain, is directly or indirectly immobilized such that the probes are oriented to have a free 3′ end to enable said probe to function as a reverse transcriptase (RT) primer; (b) contacting said substrate with a tissue sample and allowing RNA of the tissue sample to hybridize to the capture probes; (c) generating cDNA molecules from the captured RNA molecules using said capture probes as RT primers; (d) labelling the cDNA molecules generated in step (c), wherein said labelling step may be contemporaneous with, or subsequent to, said generating step; (e) detecting a signal from the labelled cDNA molecules; and optionally (f) imaging the tissue sample, wherein the tissue sample is imaged before or after step (c).

Abstract translation: 本发明涉及用于组织样本或其部分中RNA的局部或空间检测和/或分析的方法和产品，其包括：（a）提供对象底物，其上至少一种捕获探针，其包含捕获 结构域直接或间接固定，使得探针定向为具有游离的3'末端以使所述探针用作逆转录酶（RT）引物; （b）使所述底物与组织样品接触并允许组织样品的RNA与捕获探针杂交; （c）使用所述捕获探针作为RT引物从捕获的RNA分子产生cDNA分子; （d）标记步骤（c）中产生的cDNA分子，其中所述标记步骤可以与所述生成步骤同时或之后; （e）检测来自标记的cDNA分子的信号; 以及任选地（f）对所述组织样品进行成像，其中所述组织样品在步骤（c）之前或之后成像。