GSM1682610: wt20_RNA-Seq; Mus musculus; RNA-Seq
Smc3-flox/flox MEF (mouse embryonic fibroblast) cells was originally derived from European conditional mouse mutagenesis program (White et al., 2013) (http://www.informatics.jax.org/allele/MGI:4434007). The adenovirus containing Cre and GFP was purchased from Hanbio biotechnology Shanghai. MEF cells were cultured with 10% PBS in DMEM (Life technology). To avoid loss of viability in Smc3-deficient cells when they enter mitosis, we infected the cells at G0/1 stage of the cell cycle. The medium was changed two days after complete confluence. 10e9 pfu adenovirus was added to the medium of 10 cm dish for 8 hr. To allow cells to recover from viral infection, we changed the medium into serum-free DMEM and kept cells for 6 days at high confluence. MEF cells were then synchronized by dexamethasone (Sigma) with the final concentration of 100 nM for 1 hour. The cells were rinsed with PBS and cultured with serum-free DMEM. Wild type and Smc3 KO MEF cells were collected at 20, 24, 28, 32, 36, and 40 hr after synchronization. Total RNA in ZT20 and ZT32 samples were extracted. RNA-seq libraries were prepared by using Illumina TruSeq RNA Sample Prep Kit V2 and were subjected to deep sequencing with 100 bp read on HiSeq 2000 at CAS-MPG Partner Institute for Computational Biology Omics Core, Shanghai, China.
GEO Accession
GSM1682610