title: GSM8238721: Cerebellum, P0, female, F1r, GLORI; Mus musculus; OTHER
library_name: GSM8238721
library_construction_protocol: Tissues were collected and separately homogenized in the presence of TRNzol Universal reagent (TianGen, #DP424).Total RNA was extracted in Trizol reagent following the manufacturer's protocol. Poly(A)+ RNA was then isolated using Dynabeads mRNA Purification Kit (Invitrogen, #61006). Glyoxal and nitrite-mediated deamination treatment were conducted on mRNA samples. For library construction, 50 ng of the GLORI-treated RNA sample was firstly repaired and ligated to pre-adenylated 3′-adapter. After the redundant adapter was digested by Lambda Exonuclease (NEB, #M0262S) and 5′ Deadenylase (NEB, #M0331S), the RNA samples were ligated to a pre-heated 5′-adapter without cleanup. Reverse transcription was then carried out using HiScript III 1st Strand cDNA Synthesis Kit (Vazyme, #R312-01) and RT primer. Finally, the library was amplified using Q5 High-Fidelity DNA Polymerases with GC enhancer (NEB, #M0491S) and subjected to size selection using a 6% polyacrylamide gel, retaining fragments longer than 150 nt.