Multiplexed Signal Amplified FISH via Splinted Ligation Amplification and Sequencing

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...

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Bibliographische Detailangaben
Hauptverfasser: Boyden, Edward Stuart, Payne, Andrew, Alon, Shahar, Sinha, Anubhav, Wassie, Asmamaw T, Chen, Fei, Marblestone, Adam Henry
Format: Patent
Sprache:eng
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Beschreibung
Zusammenfassung: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.