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AssigneeLeica Biosystems Newcastle Ltd.
InventorsMATS NILSSON BERNITZ, Chatarina Larsson, Ida Grundberg
Estimating the term…
The present invention relates to the detection of RNA in a sample of cells. More particularly, the present invention relates to the localized detection of RNA in situ. The method relies on the conversion of RNA to complementary DNA prior to the targeting of the cDNA with a padlock probe(s). The hybridization of the padlock probe(s) relies on the nucleotide sequence of the cDNA which is derived from the corresponding nucleotide sequence of the target RNA. Rolling circle amplification of the subsequently circularized padlock probe produces a rolling circle product which may be detected. Advantageously, this allows the RNA to be detected in situ.
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The method of claim 74, wherein generating the cDNA complementary to the RNA in the sample comprises contacting said sample with a reverse transcriptase and a reverse transcription primer.
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The method of claim 74, comprising adding a ribonuclease to the sample.
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The method of claim 74, wherein the collection of padlock probes comprise different detection probe binding regions.
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A collection of padlock probes specific for mutations to the KRAS gene, comprising:
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Dependencies are inferred from the claim text.