characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-01-31T00:19:37Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-01-31T00:19:37Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110137886
characteristics.characteristic: External Id
characteristics.text: SHANGHAI JIAO TONG UNIVERSITY
characteristics.characteristic: INSDC center name
characteristics.text: 2023-01-31T00:19:37Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-01-31T00:19:37Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12235766
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11778:Bmi1_KD_SPC_H2AK119ub1_rep1
characteristics.characteristic: Submitter Id
characteristics.text: 6
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: spermatogonium
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Isolation and culture of undifferentiated spermatogonia followed the protocols with little modifications. Briefly, testis was cut off from 6 day-old pups and dissociated by a two-step enzymatic digestion. About 20 mice were used to isolate wild undifferentiated cells, and about 40 mice were used to isolate undifferentiated cells then knocked down Bmi1 by siRNA. Cells were resuspended in IMDM/FBS medium and plated onto dishes coated by 0.2% (w/v) gelatin. An hour later, the suspended cells were transferred to a new dish coated by gelatin. This step was repeated twice, then incubated overnight. Somatic cells were adhered in dishes, undifferentiated spermatogonia were loosely attached. 4x10^6 cells were used for each ChIP-seq experiment. Cells were cross-linked using a final concentration of 1% formaldehyde (Thermo Fisher, USA) at room temperature for 10 min, then stopped by a final concentration of 125mM Glycine (Sangon, China) at room temperature for 5min, subsequently washed twice with PBS. Cells were isolated nucleus according to previously described. Chromatin was sheared by Covaris sonicator (Cavaris, USA). The fragmented chromatin was immunoprecipitated with Magna ChIPTM Protein A+G Magnetic beads (Millipore, USA) coupled with antibodies overnight at 4°C. Then, the beads were washed with low salt, high salt, LiCl and TE buffer. Bound chromatin was eluted and reverse crosslinked. ChIP and input DNA were constructed libraries using KAPA Hyper Prep Kit (KAPA, USA) according to the manufacturer’s instructions
characteristics.characteristic: description
characteristics.text: infant
characteristics.characteristic: developmental stage
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: testis
characteristics.characteristic: organism part
characteristics.text: Bmi1 deficient
characteristics.characteristic: phenotype
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: ICR
characteristics.characteristic: strain
characteristics.text: Bmi1_KD_SPC_H2AK119ub1_rep1
characteristics.characteristic: title