title: GSM8017517: Astrocytes, OGD/R group, biol rep 1; Rattus norvegicus; RNA-Seq
library_construction_protocol: Total RNA was extracted from cells using TRIzol reagent according to the manufacturer's protocol. 1 μg of total RNA was subjected to library synthesis using the TruSeq V2 RNA-Seq kit.1 μg of total RNA was enriched for the Poly-A mRNA and reverse transcribed to double-stranded cDNA. The cDNA was end repaired, adenylated, and ligated with Illumina indexes prior to 12 cycles of PCR. Sequencing was performed using 12pM hybridization to a 2x100 paired end flow cell. After extracting total RNA from the sample and passing quality inspection, enrich the eukaryotic mRNA using magnetic beads with Oligo(dT) (if it is prokaryotic, use a kit to remove rRNA). Add fragmentation reagents and use a thermocycler to fragment the mRNA into short fragments. Synthesize first-strand cDNA using the fragmented mRNA as a template, and then prepare a reaction system for the synthesis of second-strand cDNA. Purify, repair sticky ends, and add a base "A" to the 3' end of the cDNA, followed by adapter ligation using a kit. Perform fragment size selection and PCR amplification to construct the library. After the constructed library passes quality inspection using the Agilent 2100 Bioanalyzer and Qubit, perform sequencing using the Illumina sequencer.