GSM6127875: Cela_1; Mus musculus; RNA-Seq
The total RNA from the ischemic hemisphere or cells was isolated with the TRIzol Reagent according to the manufacturer's instructions. A total of 5 µg RNA from ischemic hemisphere was utilized for each RNA sample. Firstly, ribosomal RNA (rRNA) was depleted by Ribo-Zero Gold rRNA Removal Kit (Illumina, USA). Secondly, the left RNAs were fragmented into short fragments using divalent cations (NEBNext® Magnesium RNA Fragmentation Module, NEB, USA) under high temperature. The complementary DNA (cDNA) was synthesized and purified. Finally, the average insert size for the final cDNA library was 300 ± 50 bp. 2×150 bp paired-end sequencing was performed on Illumina Novaseq™6000 (LC-Bio Technology Co. Ltd., China) according to the recommended protocol.