Homo sapiens
Protocols: Peripheral blood mononucleated cells (PBMC) were obtained from healthy donor buffy coats (Etablissement Français du sang, Rungis, France), separated on Pancoll (Pan-Biotech, Dutscher, Brumath, France). Peripheral blood CD14+ monocytes were sorted with magnetic beads and the AutoMacs system (Miltenyi Biotech, Paris, France) After sorting, monocytes were cultured overnight at 1 million cells/mL in RPMI 1640 Glutamax medium (ThermoFisher Scientific) supplemented with 10% heat inactivated fetal bovine serum (FBS, Lonza, Amboise, France), 1% penicillin/streptomycin and 2mM L-Glutamine (ThermoFisher Scientific). Total RNA was isolated from primary monocytes or macrophages with RNeasy mini Kit (Qiagen). The RNA integrity (RNA Integrity Score ≥ 7.0) was checked on the Agilent 2100 Bioanalyzer (Agilent) and quantity was determined using Qubit (Invitrogen). SureSelect Automated Strand Specific RNA Library Preparation Kit was used according to manufacturer's instructions with the Bravo Platform. Briefly, 100 ng of total RNA sample was used for poly-A mRNA selection using oligo(dT) beads and subjected to thermal mRNA fragmentation. The fragmented mRNA samples were subjected to cDNA synthesis and were further converted into double stranded DNA using the reagents supplied in the kit, and the resulting dsDNA was used for library preparation. The final libraries were processed using Illumina TruSeq Stranded mRNA Library Prep Kit.
ENA first public
2019-12-13
ENA last update
2017-12-13
External Id
SAMEA104428875
INSDC center alias
Inserm U1170
INSDC center name
Inserm U1170
INSDC first public
2019-12-13T04:02:40Z
INSDC last update
2017-12-13T16:41:21Z
INSDC status
public
Submitter Id
E-MTAB-6306:Donor1/BC212 Mono
age
59
broker name
ArrayExpress
cell type
monocyte
common name
human
disease
normal
growth condition
overnight in RPMI 10 percent FBS
organism part
blood
phenotype
CD14 positive
sample name
E-MTAB-6306:Donor1/BC212 Mono
sex
male
time
0