Methods and product for optimising localised or spatial detection of gene expression in a tissue sample
First Claim
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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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).
174 Citations
43 Claims
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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:
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(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). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43)
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Specification