2024-03-21
ENA first public
ERC000011
ENA-CHECKLIST
Centre for Discovery Brain Sciences, University of Edinburgh
INSDC center name
public
INSDC status
ERS18536108
SRA accession
E-MTAB-13936:TAN2-74
Submitter Id
18
month
age
ArrayExpress
broker name
astrocyte
cell_type
not collected
collection date
house mouse
common name
Protocols: Tissue was lysed in ice-cold lysis buffer (in mM: 20 HEPES, 10 MgCl2, 150 KCl, 0.5 DTT) along with 100 μg/mL cycloheximide (Sigma), cOmplete ULTRA protease inhibitors (Sigma) and RNAse inhibitors (Superasin – Life Technologies; RNAsin – Promega). Cell lysate was centrifuged to clear debris and solubilised with 1% NP40 and 30 mM 1,2-diheptanoyl-sn- glycero-3-phosphocholine (DHPC – Avanti Polar Lipids).  Following further centrifugation, supernatants were added to pre-prepared anti-GFP (19C8 and 19F7 antibodies – Sloan Kettering Memorial Centre) coated magnetic beads (Dynabeads MyOne Streptavadin T1 – Life Technologies). Following immunoprecipitation overnight at 40C with continuous rotation, beads were washed four times. Cell-type specific mRNA, attached to immunoprecipitated GFP-tagged ribosomes, was isolated and purified using the Agilent Nanoprep kit. TRAP-sequencing was performed utilising the Clonetech - SMART-Seq v4 Ultra Low input RNA library preparation.
description
adult
developmental stage
APP NL-F x Aldh1l1-EGFP-Rpl10a
genotype
not collected
geographic location (country and/or sea)
TAN2-74
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
cerebral cortex
organism part
Mus musculus
scientific_name
male
sex
C57BL/6J
strain
TAN2-74
title