GSM9081259: hippocampus-1, PXN KO,PXN-S119A KI, P15; Mus musculus; RNA-Seq
Total RNA was extracted utilizing the RNeasy Mini Kit (Qiagen; Cat# 74104) and its concentration was determined using a Nanodrop 2000 spectrophotometer and a Qubit fluorometer (both from ThermoFisher Scientific). The quality of the extracted RNA was evaluated using a 2100 Bioanalyzer (Agilent). 120 ng total RNA per sample was subjected to cDNA synthesis using SMART-Seq  mRNA with UMI (Takara,Cat. 634763) following manufacturer's instruction. Samples were amplified for 8 PCR cycles and using  0.5 x AMPure select above 500bp cDNA. The full-length cDNAs weres prepared for long-reads  sequencing using the Oxford Nanopore Ligation Native barcoding Kit 24 V14 ( SQK-NBD114.24) .barcoded cDNA library was loaded onto a PromethION flow cell (pore version 10.4.1, flow cell version FLO-PRO114M) and run on a PromethION P2 solo sequencing device for 72 hours.