2024-10-08
ENA first public
ERC000011
ENA-CHECKLIST
INSERM Gustave Roussy
INSDC center name
public
INSDC status
ERS20128847
SRA accession
E-MTAB-14124:Foetal_Liver_CTRL_noSCFnoIL3_1
Submitter Id
ArrayExpress
broker name
not collected
collection date
human
common name
Protocols: 5x104-1x105 GFP+ live-sorted cells were spun at 500g for 5 min, washed with cold DPBS, lysed in cold lysis buffer (10 mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, and 0.1% NP40, 3min on ice) and immediately spun at 500g for 10 min. The pelleted nuclei were resuspended in 50 µL of transposase reaction mix (Tagment DNA TDE1 Enzyme and Buffer Kit, Illumina) for 30 min at 39°C.  Transposed DNA was purified using a MinElute PCR purification kit  (Qiagen) in 15µL of nuclease-free H2O and amplified with NEBNext® Ultra™ II Q5® Master Mix and Nextera XT DNA Library Preparation Kit (Illumina), using the following PCR conditions: 72°C for 5 min, 98°C for 30s, then 12 cycles of 98°C for 10 s, 63°C for 30 s, and 72°C for 1 min.  Transposed DNA was purified using a MinElute PCR purification kit  (Qiagen) in 15µL of nuclease-free H2O and amplified with NEBNext® Ultra™ II Q5® Master Mix and Nextera XT DNA Library Preparation Kit (Illumina), using the following PCR conditions: 72°C for 5 min, 98°C for 30s, then 12 cycles of 98°C for 10 s, 63°C for 30 s, and 72°C for 1 min. Libraries were purified with AMPure XP beads (Beckman Coulter)
description
foetus
developmental stage
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
liver
organism part
Homo sapiens
scientific_name
No SCF No IL3
stimulus
Foetal_Liver_CTRL_noSCFnoIL3_1
title