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1. (WO2018136856) MULTIPLEXED SIGNAL AMPLIFIED FISH VIA SPLINTED LIGATION AMPLIFICATION AND SEQUENCING

Pub. No.:    WO/2018/136856    International Application No.:    PCT/US2018/014629
Publication Date: Fri Jul 27 01:59:59 CEST 2018 International Filing Date: Tue Jan 23 00:59:59 CET 2018
IPC: C12Q 1/6841
Applicants: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Inventors: CHEN, Fei
WASSIE, Asmamaw, T.
ALON, Shahar
MARBLESTONE, Henry, Adam
SINHA, Anubhav
PAYNE, Andrew
BOYDEN, Edward, Stuart
Title: MULTIPLEXED SIGNAL AMPLIFIED FISH VIA SPLINTED LIGATION AMPLIFICATION AND SEQUENCING
Abstract:
The present invention relates to a method for amplifying at least one target RNA in a fixed and, optionally, expanded biological sample. In an embodiment of the invention, the method comprises incubating the fixed biological sample with a pair of polynucleotides complementary to non-overlapping and proximal sequences of a target RNA, wherein the polynucleotide pair hybridizes to the target RNA; ligating the polynucleotide pair using a ligase; and amplifying the ligation product. The invention further provides methods for detecting and optionally quantifying and/or sequencing the amplification product. As the method comprises hybridizing polynucleotide pairs to a target RNA in a fixed biological sample, the target RNA can be hybridized in situ.