2024-04-29
ENA first public
ERC000011
ENA-CHECKLIST
Wellcome Sanger Institute
INSDC center name
public
INSDC status
ERS19298786
SRA accession
E-MTAB-8813:cell_suspension_22
Submitter Id
6.1
week
age
ArrayExpress
broker name
not collected
collection date
human
common name
Protocols: The human embryonic samples were collected from medical abortions. The samples were stored in hibernate E medium (Thermo Fisher A1370601) and transported to the research lab for dissection. The whole limb bud or upper part of the lower limbs were then dissected are stored in ice-cold PBS before single-cell isolation. Samples were chopped into a mash using a scalpel and incubated in 5ml of 2.5mg/ml liberase at 37 degrees for 30-40 min till fully digestion. Cells were spun down at 4C 750 for 5min and resuspended in 3ml of 1X RBC lysis buffer (eBioscience 00-4333, or diluted from eBioscience 00-4300) followed by 4-5 min incubation at RT. 10ml of 1X PBS was then added. The cells were then spun down at 4C at 750 for 5min, resuspended in 5ml of 2% FBS in 1X PBS, filtered through a 40um strainer, spun down again iat 4C 750g for 5min, resuspended in 50-200ul of 2% FBS in PBS, and spun down again. Libraries were prepared according to the standard protocol of the Chromium single cell 3’ V2 kit
description
embryo stage
developmental stage
normal
disease
not collected
geographic location (country and/or sea)
BRC2243
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
forelimb
organism part
sampling site
Homo sapiens
scientific_name
male
sex
cell_suspension_22
title