2025-02-28
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA113400436
External Id
DKFZ Heidelberg
INSDC center name
2023-05-03T13:48:23Z
INSDC last update
public
INSDC status
ERS15395581
SRA accession
E-MTAB-12899:Sample 12
Submitter Id
ArrayExpress
broker name
pancreatic cell
cell_type
human
common name
Protocols: Ganglia cells were seeded as described above and after axons started to grow out, cells were incubated for 1 hour with cell-trace violet (ThermoFisher C34557) to enable labelling. Cells were washed several times, fresh medium was added and PACO10-GFP (Sigma Aldrich MISSION® pLKO.1-puro-CMV-TurboGFP™ Positive Control Plasmid DNA) cells were added to the well. The cells were cultured together for 3 days before FACS sorting of PACO cells. For cell proliferation, ganglia cells were seeded as described above and PACO/fibroblast were cultured in a transwell chamber. RNA extraction and purification of FACS-sorted cells was done using PicoPure RNA Isolation Kit according to manufacturer’s instructions (Thermo Fisher, cat # KIT0214). RNA quality assessment and quantification were performed with Bioanalyzer using Agilent RNA 6000 Pico Kit (Agilent, cat # 5067-1513). Whole transcriptome amplification was performed using a modified smart-seq2 protocol [49], with 5μl of a modified RT buffer containing 1× SMART First Strand Buffer (Takara Bio Clontech, cat # 639538), 1 mM dithiothreitol (Takara Bio Clontech), 1 μM template switching oligo (IDT), 10 U μl−1 SMARTScribe (Takara Bio Clontech, cat # 639538) and 1 U μl−1 RNasin Plus RNase Inhibitor (Promega, cat # N2615). Tagmentation of cDNA was done using Nextera XT DNA Library Preparation Kit (Illumina, cat # FC-121-1030).
description
adult
developmental stage
pancreatic ductal adenocarcinoma
disease
wild type genotype
genotype
coculture with murine dorsal root ganglia
growth condition
Lane2_ASID
individual
Homo sapiens
organism
Homo sapiens
organism
pancreas
organism part
15
sampleid
Homo sapiens
scientific_name
Sample 12
title