2025-08-09
ENA first public
ERC000011
ENA-CHECKLIST
1 Institute of Human Biology (IHB), Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel, Switzerland 2 Institute of Molecular and Clinical Ophthalmology Basel, Basel, Switzerland
INSDC center name
public
INSDC status
ERS24421846
SRA accession
E-MTAB-15107:Spheroids_H9-WT_D4_S115
Submitter Id
ArrayExpress
broker name
H9-WT
cell line
midgut/hindgut patterned spheroids
cell type
not collected
collection date
human
common name
Protocols: Human and chimpanzee spheroids budding at day 3 to day 6 of midgut/hindgut patterning were sorted manually into a 96-well format. Spheroids were lysed in a lysis buffer according to the original protocol of FLASH-seq. Reverse transcription and PCR amplification were carried out using SuperScript IV Reverse Transcriptase ( Thermo Fisher, 18090200) and KAPA HiFi HotStart ReadyMix (Roche, 07958935001). cDNA was purified using SeraMag SpeedBeads (Cytiva, 65152105050250) (0.8:1 bead-to-cDNA ratio) and quantified using the Quant-iT PicoGreen dsDNA assay (Thermo Fisher, P7589). Size distribution was assessed via an Agilent 2100 Bioanalyzer. Samples were normalized to 200 pg/µl for downstream library preparation.  Tagmentation was performed using homemade Tn5 transposase followed by heat inactivation with SDS. Enrichment PCR was conducted with KAPA HiFi DNA Polymerase (Roche, KK2502) and index adapters.
description
budding at day4 of midgut/hindgut patterning
developmental stage
normal
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
midgut/hindgut
organism part
Homo sapiens
scientific_name
Spheroids_H9-WT_D4_S115
title