2024-11-19
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114320962
External Id
Tel Aviv University
INSDC center name
2023-09-01T11:52:18Z
INSDC last update
public
INSDC status
ERS16310074
SRA accession
E-MTAB-13276:bat5_GEX
Submitter Id
ArrayExpress
broker name
not collected
collection date
Egyptian rousette
common name
Protocols: Lung tissues of 8 bats were removed following transcardial perfusion and washed in cold D-PBS (Biological Industries, 02-023-1A). Trachea was removed and the tissue was immediately injected with an enzyme mix based on previous work [A cellular census of human lungs identifies novel cell states in health and in asthma] (Dispase II 50 caseinolytic u/ml (Sigma-Aldrich, D4693), Elastase 4.3 u/ml (Worthington Biochem, Worthington, LS002292)), Dnase I 30 μg/ml (Roche), Collagenase A 2 mg/ml (Roche, 10103578001), CaCl2 5 mM) followed by mincing and a brief incubation in a CO2 incubator at 37 ºC. Samples were then transferred to a shaker-incubator for 20 min at 37 ºC, 150 rpm, followed by an immediate addition of 15 ml of neutralization buffer (DMEM (Biological industries, 01-052-1A), 10% FBS (Rhenium, 10270106). Samples were then passed through a 40 um strainer twice and pelleted at 400 G, 5 min at 4 °C. Samples were then treated with RBC cell lysis (Sigma, R7757 as according to the manufacturer’s protocol) and quenched with (10% FBS, D-PBS). Cells were then pelleted at 400 G, 5 min at 4 °C, resuspended in DMEM and counted. Dead cell removal protocol (Miltney, 130-090-101 ) was applied according to manufacturer protocol in cases when cell viability was lower than 75%, and the flowthrough was pelleted at 400 G ,5 min at 4 °C and resuspended in PBS with 0.05% BSA. Cells were counted and loaded on the 10X Genomics Chromium instrument for single-cell sequencing, using 3’ V2 chemistry, according to the manufacturer’s protocol. Cells were counted and loaded on the 10X Genomics Chromium instrument for single-cell sequencing, using 3’ V3 chemistry, according to the manufacturer’s protocol. cDNA library preparation was were carried out according to the manufacturer’s protocol.
description
adult
developmental stage
normal
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
bat5
individual
not applicable
isolate
Rousettus aegyptiacus
organism
Rousettus aegyptiacus
organism
lung
organism part
Rousettus aegyptiacus
scientific_name
male
sex
bat5_GEX
title