D_KO_R3
Mus musculus
Protocols: Hippocampi were dissected from postnatal day 1 (P1) C57/BL6, KD-/-, K-/-, or D-/- mice and mechanically dissociated by fine mincing and enzymatically digested for 30 minutes at 37ºC in DMEM media containing 2.5U/ml Papain, 1U/ml Dispase II , and 250U/ml DNase I. NSCs were purified using a 65% Percoll gradient and cultured (Neural Basal A medium supplemented with 2% B27 without Vitamin A, 1% Glutamax, 1% Penicillin Streptomycin, 10ng/ml EGF, 10ng/ml bFGF) as a monolayer on poly-D-lysine and laminin-coated plates at a density of 105 cells/cm2. mRNA of NSCs was isolated by lysis with TRIZOL Reagent (Thermo Fisher Scientific), separation with chloroform (0.2 mL per mL TRIZOL), and precipitated with isopropyl alcohol. Libraries were prepared using the Illumina Nextera XT DNA Sample Preparation Kit according to the manufacturers protocol. cDNA was validated using KAPA qPCR-based library quantification kit.
ENA first public
2018-01-01
ENA last update
2017-06-19
External Id
SAMEA104128080
INSDC center alias
Dept. of Anatomy UCSF
INSDC center name
Dept. of Anatomy UCSF
INSDC first public
2018-01-01T17:01:41Z
INSDC last update
2017-06-19T08:35:23Z
INSDC status
public
Submitter Id
E-MTAB-5827:D_KO_R3
age
1
broker name
ArrayExpress
cell type
neural stem cell
common name
house mouse
developmental stage
neonate
genotype
H2-D1 knockout
organism part
hippocampus proper
sample name
E-MTAB-5827:D_KO_R3
strain
C57BL/6