title: GSM8498205: ZAR1_PAIso-seq2; Mus musculus; RNA-Seq
library_construction_protocol: 100 GV and MII stage oocytes harvested from WT and Zar1/2-/- mice and 100 MI stage oocytes harvested from WT mice were put into PAIso-seq2 library construction as published protocol. In brief, total RNA was extracted with TRIzol Reagent and Direct-zol RNA MicroPrep (Zymo Research, Cat. no. R2060), and the barcode containing adapter were ligated to the 3'-end of mRNA. Then the RNA were purified by the RNA Clean & Concentrator-5 kit (Zymo Research, Cat. no. R1016). After being purified by the RNA Clean & Concentrator-5 kit (Zymo Research, Cat. no. R1016), the RNA were RT into cDNA by the UMI containing template-switching oligo (TSO) primer and the first PCR amplification were proceeded. Then the cDNA from rRNA will be removed by the CRISPR-Cas9 system and the second PCR amplification will be performed to get enough cDNA. Then the cDNA from each sample will be concatenated into long molecules by following the MAS-ISO-seq procedure. Finally, all the samples were mixed up according to the molar ratios of cDNA, and at least 500ng of cDNA was sent for SMRTbell library preparation and sequenced on PacBio Revio platform.