Patent ID: 11859234
Assignee: THE TRUSTEES OF PRINCETON UNIVERSITY
Field: Measurement (Instruments)
Classification: CPC C  A  G | IPC A  C  G

Claim 0:
1. A method for detecting an RNase L cleavage product comprising,
a) isolating total RNA or small RNAs from a sample;
b) ligating a ligation adapter polynucleotide to the RNA from the sample cleaved by RNase L, wherein the ligation adapter polynucleotide specifically ligates to RNA that has been cleaved by RNaseL, wherein the ligation adapter polynucleotide has at its 5′ end at least 1 nucleotide that is identical to a 3′ end of a first primer;
c) generating cDNA by reverse-transcription of the RNA-ligation adapter polynucleotide ligation product using a reverse transcription primer that is complementary to at least a portion of the ligated ligation adapter polynucleotide;
d) contacting the cDNA with the first primer, having at its 3′ end at least 1 nucleotide identical to the ligation adapter polynucleotide 5′ end and further comprising nucleotides identical to at least a portion of the sequence of RNase L-cleaved RNA and having one or more bases identical to the ligation adapter polynucleotide but not to uncleaved isolated RNA, and a second primer that is identical to at least a portion of the reverse transcription primer;
e) amplifying the cDNA encoding the RNA-ligation adapter polynucleotide product; and
f) detecting the amplified product of (e) to detect a RNase L cleaved RNA product (“RNase L cleavage product”).