2025-02-28
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA115112563
External Id
Prinses Maxima Centrum voor Kinderoncologie
INSDC center name
2024-01-17T19:12:58Z
INSDC last update
public
INSDC status
ERS17748305
SRA accession
E-MTAB-13715:E_18_NC
Submitter Id
ArrayExpress
broker name
not collected
collection date
human
common name
Protocols: For RNA isolations 2 wells were washed briefly with icecold PBS and cells were lysed directly in TRIzol (ThermoFisher Scientific 15596026 ). For Immunoblotting 2 wells were washed with PBS and cells were lysed directly in 2xLaemmli sample buffer with β-mercaptoethanol. RMS000EEC organoids were cultured as described in Meister et al. (55) Briefly early passage cryovials were restarted at high density (50,000 cells/cm2) in BM1* medium supplemented with 0.3% BME. Cells were growing at 37o C and 5% CO2 as attached cells with a doublingtime of 144 hrs, and passaged to start from 25,000 cells/cm2. siRNA transfections were performed by harvesting RMS000EEC cells and replating in 24 well plates at 200,000 cells per well in 500ul BM1* with 0.3% BME. After 24 hours transfections were done by adding to each well a 50ul mixture of Lipofectamine RNAiMAX (ThermoFisher Scientific 13778100) and DsiRNA in Optimem (ThermoFisher Scientific 31985062), such that the endconcentration in the well is 3ul LF/ml, 30nM DsiRNA . Cells were washed after 24 hours and harvested after 48 hours. NEBNext® UltraTM RNA Library Prep Kit (New England Biolabs) was used, according to manufacturer’s instructions, to prepare the sequencing libraries. Strand specific mRNA libraries were prepared by Novogene (UK). RNA samples were sequenced using a paired-end approach
description
embryonal rhabdomyosarcoma scrambled
disease
not collected
geographic location (country and/or sea)
organoid culture
growth condition
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
2
replicate
Homo sapiens
scientific_name
E_18_NC
title