2025-05-12
ENA first public
ERC000011
ENA-CHECKLIST
EMBL
INSDC center name
public
INSDC status
ERS20058536
SRA accession
E-MTAB-14107:GFP2_ATAC_L3
Submitter Id
ArrayExpress
broker name
not collected
collection date
fruit fly
common name
Protocols: We used 20 imaginal wing discs per sample to make two technical replicates, and the larvae were of salEPV-Gal4 UAS-salm-i/+; UAS-salr-i/+ and salEPV-Gal4 UAS-GFP/+ genotypes. The discs were dissected in pre-cooled PBS, pipetted in PBT (PBS, 0.1 % Triton X-100) containing 1x proteases inhibitors (cOmplete™ Protease Inhibitor Cocktail tablets, Roche, 11873580001) and suspended in transposition solution containing. Disk were permeabilization: Thaw disks on ice for 10 min and add 50ul PBT+PI; Add 50 ul of PBT + 0.4% IGEPAL CA-630; Incubate at 4C for 30 min on a nutator and Spin at 3200 xg for 5 min at 4C. Tagmentation and reverse-crosslinking: Resuspend the nuclei pellet in 50 ul tagmentation mix(5 uL TDE1 (Illumina, 15027865), 25 uL 2x buffer (Illumina, 15027866), 20 uL nuclease-free H2O). Incubate at 37C for 30 min. Add 50 ul STOP buffer, 5 ul of 1mg/ml RNase A and incubate at 55C for 10 min. Add 3 ul Proteinase K (20 mg/ml) incubate 65C for 1 hour. Purify with Zymo DNA clean and concentrator kit. Finally samples are PCR amplified and size selected with SPRI beads.
description
L3;Third Instar
developmental stage
transgenic
genetic modification
SalEPV-Gal4 /UAS-GFP
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Drosophila melanogaster
organism
Drosophila melanogaster
organism
Wing imaginal disc
organism part
frozen_sample
sample type
Drosophila melanogaster
scientific_name
SalEPV-Gal4 /UAS-GFP
strain
GFP2_ATAC_L3
title