title: GSM6253066: Con-2; Mus musculus; RNA-Seq
library_construction_protocol: Cloning of SERPINE2 gene CDs, double enzyme digestion, TOS189-1 vector double enzyme digestion, ligation, transformation, bacterial solution PCR and sequencing comparison, the lentivirus vector was packaged and constructed by Starfish Biotechnology (Suzhou) Co., Ltd. Consistently, control virus—pLV[Exp]-EGFP:T2A:Puro-EF1A>mCherry also provided by the company. GCs were transfected with a negative control duplex or siRNAs against SERPINE2 using Lipofectamine 3000 Transfection Reagent (Thermo Fisher Scientific) following the manufacturer's instructions. After screening the multiplicity infection, lentivirus was transfected into GCs following the manufacturer's instructions and the positive cells were screened by puromycin dihydrochloride. 48 h after transfection, the cells were collected for RNA sequencing.Total RNA was extracted from 1Í106 cells using TRIzol Reagent (Thermo Fisher Scientific). 3 mg per sample were constructed into libraries through TruSeq Stranded mRNA LTSample Prep Kit for Illumina (RS-122-2101) following manufacturer's manuals. Sequencing data were generated on Illumina HiSeqTM 2500 platform with 150-bp paired-end read length by OE Biotech Co., Ltd (Shanghai, China). RNA libraries were prepared for sequencing using standard Illumina protocols