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: SAMEA110138384
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: ERS12237204
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11777:Bmi1 KD SPC-rep2
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 in wild and Bmi1 knockdown group. Total RNA was isolated by Trizol (Invitrogen) according to the manufacturer’s protocols. mRNA was purified by NEBNext Poly(A) mRNA Magnetic Isolation Beads (NEB, USA). The RNA libraries were constructed using the NEBNext Ultra Directional RNA Library PrepKit (NEB, USA).
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-rep2
characteristics.characteristic: title