transgenic carrying the tetO-OSKM transgene and Oct4-GFP/Rosa26-M2rtTA
cell type
reprogrammable mouse embryonic fibroblasts (MEFs)
induced with
doxycycline (Dox) (1ug/ml) for 3 days
reprogramming stage
day3
dox-induced genes
OSKM+Obox1
chip antibody
none
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
For RNA-seq, the cells were lysed using RLT buffer supplemented with β-mercaptoethanol and processed using the RNeasy mini kit (Cat. 74104).For ChIP-seq, the cells were cross-linked with 37% formaldehyde in serum-free medium at room temperature for 10 min in room temperature (RT). Glycine was added to stop the cross-linking at final concentration of 0.125 M and the cells were rinsed in PBS. Then the cells were re-suspended in lysis buffer containing protease inhibitors and chromatin was sonicated to 100-400 bp with Covaris M220 system. RNA-seq: KAPA Stranded mRNA-Seq Kit (KK8420) ChIP-seq: KAPA HyperPlus Library Preparation kit (KAPA KK8510)