title: K_KO_R1
organism: Mus musculus
description: 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.
attributes.tag: ENA first public
attributes.value: 2018-01-01
attributes.tag: ENA last update
attributes.value: 2017-06-19
attributes.tag: External Id
attributes.value: SAMEA104128081
attributes.tag: INSDC center alias
attributes.value: Dept. of Anatomy UCSF
attributes.tag: INSDC center name
attributes.value: Dept. of Anatomy UCSF
attributes.tag: INSDC first public
attributes.value: 2018-01-01T17:01:41Z
attributes.tag: INSDC last update
attributes.value: 2017-06-19T08:35:23Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5827:K_KO_R1
attributes.tag: age
attributes.value: 1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: neural stem cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: neonate
attributes.tag: genotype
attributes.value: H2-K1 knockout
attributes.tag: organism part
attributes.value: hippocampus proper
attributes.tag: sample name
attributes.value: E-MTAB-5827:K_KO_R1
attributes.tag: strain
attributes.value: C57BL/6