2024-05-15
ENA first public
ERC000011
ENA-CHECKLIST
Swedish University of Agricultural Science, Uppsala
INSDC center name
public
INSDC status
ERS19850106
SRA accession
E-MTAB-14078:P18856_4003_S17
Submitter Id
ArrayExpress
broker name
epithelial cell
cell_type
not collected
collection date
59.78233273
collection latitude
17.62738681
collection longitude
caught in the wild
collection site
Protocols: L. sclopetarius adult female spiders were collected in the wild in a small habitat in Uppsala, Sweden. Taxonomic identity of the spider was verified by the Museum of Natural History, Stockholm, Sweden. The spiders were kept in big containers that allowed them to spin webs. They were fed with meal worms or Drosophila flies weekly and watered daily. In total 20 spiders were used for single cell sequencing.  Method B: Due to problems with contaminating droplets of dope that made the separation of single cells challenging, a slightly modified protocol was used for the following ten spiders. These were processed as method A in Protocol 2, but with the exception that the duct was not removed, and the sac was cut and kept in PBS, pH 7.4 for 10 min to allow the dope to flow out. The pieces of glands were picked up and incubated in pre warmed trypsin-EDTA (Gibco, 0.5%) at 37°C for 2 min in a low-binding micro-centrifuge tube. The suspension was triturated with fire-polished glass Pasteur pipette for 1 min, incubated in trypsin-EDTA at 37°C for 2 min, and triturated again with fire polished glass Pasteur pipette for 3 min. The suspension was centrifuged at 300 23g for 3 min at 4°C and the pellet was resuspended in 200 μL DMEM containing 3% BSA. The suspension was strained through 40 μM pre-washed cell strainers into low-binding micro-centrifuge tubes. The strainer was further washed with 100 μL DMEM containing 3% BSA to reduce the loss of cells. The library preparation was done using 10X 3’ GE kit on the 10X Chromium Single Cell 3’ Platform following manufacturer’s protocol (Dual Index 10X_3'_V3.1).
description
adult
developmental stage
wild type genotype
genotype
Sweden
geographic location
not collected
geographic location (country and/or sea)
not applicable
individual
not applicable
isolate
Larinioides sclopetarius
organism
Larinioides sclopetarius
organism
Major ampullate gland
organism part
27
replicate
Larinioides sclopetarius
scientific_name
female
sex
Silked
silked status
not applicable
strain
P18856_4003_S17
title