GSM9369239: Cleavage site profiles (PspCas13b.T136A with crRNA targeting HTT-EGFP, cr7,  replicate 2); Homo sapiens; OTHER
RNA was extracted by RNA mini-prep (Zymo) according to manufacturer's instruction To map the 5' end of Cas13 cleavage sites on target RNA, we carried out 5' RACE and an in vivo adaptor ligation method. For 5' RACE, a template-switching oligo (TSO) was included during reverse transcription by following a previous protocol (EMBO Rep 2025, 26:1891). For the in vivo adaptor ligation library, the TSO is not required. A total of 2 rounds of PCR were performed to amplify the libraries using either a TSO-specific forward primer or an in vivo adaptor-specific forward primer, coupled with a target-gene-specific reverse primer.