c-Myc supports polyploidy and prevents senescence in the murine placenta [RNA-seq]

To understand differential gene expression between wildtype and c-Myc null mouse trophoblast giant cells. TGCs from 9.5 dpc stage were dissected as mentioned previously (Hannibal and Baker, 2016) cold PBS and embryos were used for genotyping. Total RNA from TGCs was isolated using TRIzol (#15596026, Thermo Fisher Scientific). rRNA was depleted using NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB # E6310L). cDNA synthesis was performed using NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760S) and NEBNext Multiplex Oligos for Illumina (Index Primer Set 1) (NEB # E7335S). Sequencing was performed using an Illumina NextSeq 500 with 75bp single read runs.本研究旨在探究野生型（wildtype）与c-Myc敲除（c-Myc null）小鼠滋养层巨细胞（trophoblast giant cells）之间的差异基因表达情况。采集妊娠9.5天（days post coitum, dpc）的滋养层巨细胞（简称TGCs），按照此前文献报道的方法（Hannibal与Baker, 2016）完成解剖分离；随后使用预冷磷酸盐缓冲液（phosphate buffered saline, PBS）处理胚胎以开展基因分型（genotyping）。采用TRIzol试剂（货号#15596026，赛默飞世尔科技（Thermo Fisher Scientific））提取TGCs的总RNA（total RNA）；使用NEBNext核糖体RNA（ribosomal RNA, rRNA）去除试剂盒（人/小鼠/大鼠适用，货号#E6310L，新英格兰生物实验室（NEB））完成rRNA去除步骤。采用NEBNext Ultra II定向RNA文库制备试剂盒（货号#E7760S，NEB）与NEBNext Illumina多重寡核苷酸索引引物套装1（货号#E7335S，NEB）完成互补脱氧核糖核酸（complementary DNA, cDNA）合成。使用Illumina NextSeq 500测序平台开展75bp单端测序。