organism: Homo sapiens
description: Protocols: Cells were maintained in RPMI 1640 medium (Gibco, USA) supplementary with 10% fetal bovin serum (FBS) (Gibco, USA) and 100 units/mL of penicillin and streptomycin (Gibco, USA). Cells were incubated at 37℃ in a humidified atmosphere of 5% CO2 NCI-H1975 cells were cultured in 6-well plates for 24 hours, and then were treated with Anlotinib (8 μg/mL) for 24 hours. Trizol reagent (Life Technologies, Inc., USA) was used to lyse cell samples, followed by total RNA isolation using standard procedures. mRNA was isolated using Oligotex mRNA Mini Kit (Qiagen, Germany). Residual genomic DNAs were degraded by adding DNase (NEB, USA), incubated for 30 minutes at 37℃. 100 ng mRNA from each sample was reverse-transcribed into cDNA and fragmented to 100-300 bp length. NEBNext Ultra Directional RNA Library Prep Kit (NEB, USA) was used to end-repair and adapter-ligate the fragment cDNA. Samples were amplified 10-12 cycles in a thermal cycler, in the medium of PCR Master Mix supplement with Q5 High-Fidelity DNA Polymerase (NEB, USA).
attributes.tag: ENA first public
attributes.value: 2018-11-30
attributes.tag: ENA last update
attributes.value: 2018-02-08
attributes.tag: External Id
attributes.value: SAMEA104570328
attributes.tag: INSDC center alias
attributes.value: Shanghai Chest Hospital, Shanghai Jiaotong University
attributes.tag: INSDC center name
attributes.value: Shanghai Chest Hospital, Shanghai Jiaotong University
attributes.tag: INSDC first public
attributes.value: 2018-11-30T11:01:53Z
attributes.tag: INSDC last update
attributes.value: 2018-02-08T09:35:07Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5997:Anlotinib 1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: NCI-H1975
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: non-small cell lung carcinoma
attributes.tag: organism part
attributes.value: lung
attributes.tag: sample name
attributes.value: E-MTAB-5997:Anlotinib 1