ERC000011
ENA-CHECKLIST
2022-08-14T00:13:06Z
ENA-FIRST-PUBLIC
2022-08-14T00:13:06Z
ENA-LAST-UPDATE
SAMEA10792486
External Id
European Research INstitute for the Biology of Ageing
INSDC center name
2022-08-14T00:13:06Z
INSDC first public
2022-08-14T00:13:06Z
INSDC last update
public
INSDC status
ERS8443718
SRA accession
E-MTAB-11131:GM12982_R1
Submitter Id
not available
age
ArrayExpress
broker name
lymphoblastoid cell line
cell line
lymphoblast
cell type
human
common name
Protocols: The cell lines from Bloom Syndrome and control patients were obtained from the Cell Repository of Coriell Institute: GM07492 and GM07545 (primary fibroblasts, control), GM02085 and GM03402 (primary fibroblasts, BS), GM12892 (EBV-transformed B-lymphocytes, control), GM16375 and GM17361 (EBV-transformed B-lymphocytes, BS). Mouse ES cell line F121.6 (F1 hybrid between 129/Sv and Cast/EiJ) was a kind gift from Prof. J. Gribnau (Erasmus MC, Rotterdam, The Netherlands). Blm mutant ES cell lines were generated as previously described (7). Fibroblasts were cultured in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) supplemented with 10% v/v fetal bovine serum (FBS) (Sigma Aldrich) and 1% v/v penicillin-streptomycin (Life Technologies), B-lymphocytes in RPMI1640 (Life Technologies) supplemented with 15% v/v FBS and 1% v/v penicillin-streptomycin. ES cells were cultured on mitotically arrested mouse embryonic fibroblast cells in DMEM (Life Technologies), supplemented with 15% v/v FBS (Bodinco BV), 1% v/v penicillin-streptomycin, 1% v/v non-essential amino acids (Life Technologies), 50 µM 2-mercaptoethanol (ThermoFisher Scientific), and 1,000 U ml−1 leukemia inhibitor factor (Merck). All cells were cultured at 37 °C in 5% CO2. Exponentially growing cells were harvested from individual cultures for each sample, and RNA was isolated using the Nucleospin RNA kit (Macherey Nagel). Libraries were generated for each of the three replicates, with exception of one sample where isolation produced RNA with a low RIN score (below 7) and thus was done in duplicate. RNA-sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep kit for Illumina (NEB) combined with the NEBNext rRNA Depletion kit (NEB). Complementary DNA quality and concentrations were assessed using the High Sensitivity dsDNA kit (Agilent) on the Agilent 2100 Bio-Analyzer and the Qubit 2.0 Fluorometer (Life Technologies).
description
normal
disease
Homo sapiens
organism
Homo sapiens
organism
blood
organism part
Homo sapiens
scientific_name
female
sex
GM12982_R1
title