GSM5310517: WT 24h rep1 rep2 subreads; Mus musculus; OTHER
Total RNA was extracted from cells using RNAzol Reagent. To avoid genomic DNA contamination, total RNA was treated with RQ1 RNase-Free DNase for 30 min at 37 °C, and subsequently purified using the phenol-chloroform extraction and ethanol precipitation. About 500 ng total RNA was subjected to barcoded end extension, template switching, full-length cDNA amplification, and circular adapter ligation. PAIso-seq was performed to precisely decipher the length and base composition of RNA poly(A) tails using the resultant circular full-length cDNA libraries on the PacBio third-generation sequencing platform. PAIso-seq