2024-11-19
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114295972
External Id
Tel Aviv University
INSDC center name
2023-08-22T21:35:45Z
INSDC last update
public
INSDC status
ERS16285080
SRA accession
E-MTAB-13277:bat7
Submitter Id
ArrayExpress
broker name
not collected
collection date
Egyptian rousette
common name
Protocols: Section from gut tissue sections from eight bats were collected following transcardial perfusion in cold D-PBS (02-023-1A, Biological Industries). Intestine was sectioned such that the region we took samples from included the lower part of the upper third intestinal tract and washed in cold D-PBS. Samples were cleaned and then sliced longitudinally, cleaned and again washed with D-PBS. Next, samples were minced and 0.15-0.2g of the minced tissue was transferred into the digestion mix (1.07 wU/ml Liberase DH (Roche, 5401054001), (70 U/ml) Hyaluronidase (Sigma-Aldrich 385931-25KU), (70 ug/ml) DNAse (11284932001 ,Roche) I in HBSS buffer (H6648 ,Sigma-Aldrich). The samples were incubated for 25 minutes on a shaking platform at 150rpm, and gently mixed every 10 min. Following incubation, the cells were passed through a 40mM filter into a 50ml falcon and added with 12ml of Neutralization media (DMEM, Rhenium (41965039), 10% Heat inactivated FBS (Rhenium, 10270106)). The cells were pelleted using centrifugation (400 G, 4C for 5 minutes), resuspended in DMEM and counted. Depending on red blood cell (RBC) presence in the pellet, samples were then treated with RBC cell lysis (Sigma, R7757, according to the manufacturer’s protocol), quenched with 10% FBS in D-PBS,and again pelleted and resuspended. Dead-cell removal protocol (Biotec Miltenyi, 130-090-101) was applied according to the manufacturer’s protocol in cases when cell viability was lower than 75%. The flowthrough was then pelleted at 400G and resuspended in PBS with 0.05% BSA. Cells were counted and loaded on the 10X Genomics Chromium instrument for single-cell library preparation, aiming to capture 10,000 cells, using 3’ v2 chemistry according to the manufacturer’s protocol cDNA library preparation was were carried out according to the manufacturer’s protocol.
description
adult
developmental stage
not collected
geographic location (country and/or sea)
bat7
individual
not applicable
isolate
Rousettus aegyptiacus
organism
Rousettus aegyptiacus
organism
intestine
organism part
Rousettus aegyptiacus
scientific_name
male
sex
bat7
title