| 摘要 |
The disclosure provides methods and compositions for assessing by sequencing the type and quantity of in vivo or in vitro breakdown products and stability of nucleic acids such as small interfering RNA (siRNA) and other target oligonucleotides. In general, the disclosed methods involve the use of tester oligonucleotides that comprise a double-stranded region, an optional loop (in the case of hairpin testers), and a single-stranded 3' overhang that is complementary to the full-length and a shortened target oligonucleotide. A tester is designed so that the 3' end of a respective target oligonucleotide anneals to the overhang immediately adjacent to the 5' end of the tester. The juxtaposed ends of the tester and target at adjacent positions allow for a ligase to ligate the chain if there is a match between a tester and its respective target. By sequencing the ligated product in the region of the ligation site, one may determine the sequence of the 3' end of the target oligonucleotide or of the entire target and its relative amount in the sample.
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