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Methods and product for optimising localised or spatial detection of gene expression in a tissue sample

  • US 9,593,365 B2
  • Filed: 10/16/2013
  • Issued: 03/14/2017
  • Est. Priority Date: 10/17/2012
  • Status: Active Grant
First Claim
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1. A method 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 hybridise to the capture probes under a set of conditions;

    (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).

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