GSM5128890: sRNA Retina P1; Mus musculus; ncRNA-Seq
Retinas were dissected from mice in six developmental periods: embryonic day (E) 14.5, postnatal day (P) 1, 7, 12, 17 and adult. Briefly, the mice were euthanized, and retinas were isolated from eyes and moved into the RNA-Solv Reagent (Omega Bio-tek, Norcross, GA, United States). T10 basic S25 ULTRA-TURRAX Disperser (IKA, China) was used to homogenize retinas. Total RNA was extracted by miRNA Kit (Omega Bio-tek) according to the manufacturer's instructions. RNA quantity was assessed by Agilent 2100 Bioanalyzer (Santa Clara, CA). Three RNA libraries were constructed individually to fully characterize the complex transcriptome of developing retina. Libraries of lncRNA were prepared after removed rRNA using the Ribo-ZeroTM kit (Epicentre, Madison, WI) and sequenced with pair-end 150bp using Illumina HiSeq 3000 platform. Similarly, sRNA libraries were pooled and sequenced in Illumina HiSeq 2500 sequencing platform at RiboBio (Guangzhou, China). In term of circRNA library, the rRNAs were depleted from total RNA using Ribo-ZeroTM kit. Then, RNA was treated with RNase R (Epicentre, Madison, WI) to remove liner transcripts and fragmented to approximately 200bp. The purified RNA fragments were subjected to first strand and second strand cDNA synthesis following by adaptor ligation according to instructions of NEB Next Ultra TM RNA Library Prep Kit for Illumina (NEB, USA). The purified library products were evaluated using the Agilent 2200 TapeStation and Qubit®2.0 (Life Technologies, USA) and sequenced on HiSeq 3000 with pair-end 150 bp mode.