2025-02-28
ENA first public
ERC000011
ENA-CHECKLIST
Istituto Italiano di Tecnologia - IIT
INSDC center name
public
INSDC status
ERS20877569
SRA accession
E-MTAB-14355:Sample 27
Submitter Id
ArrayExpress
broker name
NCI-H1299
cell_line
lung cancer cell
cell_type
not collected
collection date
human
common name
Protocols: Transfections were carried out in 24-well plate for flow cytometry analysis.   H1299 cells were transfected with Lipofectamine® 3000 according to manufacturer’s instructions and 300 ng of total DNA in 24-well plates. CHO and HEK293 were transfected with PEI transfection reagent with 300 ng of total DNA in 24-well plates. All cells were plated approximately 24h before transfection at 70000 to 80000 cells per well in 24-wells plates. At the moment of transfection, cells were put in starvation conditions, meaning in poor media for the first 24h. DNA was diluted in Opti-MEM reduced serum media (Gibco), before being mixed and incubated for 25 minutes prior to addition to the cells. All drugs were added 4h after transfection.  Modified-RNAs (modRNA) were transfected using Lipofectamine® 3000 protocol under modified conditions in which no P3000 reagent was used. RNA was first diluted in Opti-MEM reduced serum media (Gibco) and it was then mixed with diluted Lipofectamine reagent as manufacturer’s instructions. The mix was incubated for 7 minutes prior to addition to the cells. The fast-forward protocol was used by seeding 120000 cells per well in 24-wells plates at the moment of transfection.  Libraries were prepared using QuantSeq 3’ mRNA sequencing for RNA quantification kit (Lexogen)
description
adult
developmental stage
non-small cell lung carcinoma
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
lung
organism part
Homo sapiens
scientific_name
No Transfection
stimulus
Sample 27
title