2024-11-15
ENA first public
ERC000011
ENA-CHECKLIST
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences
INSDC center name
public
INSDC status
ERS21374133
SRA accession
E-MTAB-14565:G3BP1_NC_IP_2
Submitter Id
1
day
age
ArrayExpress
broker name
not collected
collection date
mallard
common name
Protocols: Tissues containing at least 1×107 cells were harvested and resuspended in 2 ml of nuclear separation buffer (1.28 M sucrose, 40 mM Tris pH 7.5, 20 mM MgCl2, and 4% Triton X-100) and kept on ice for at least 30 min with frequent mixing. The pelleted nuclei were centrifuged at 14,000 rpm for 15 min and resuspended in wash buffer (150 mM KCl, 25 mM Tris (pH 7.4), 5 mM EDTA, 0.5 mM DTT, 0.5% NP40, 100 U/ml RNase inhibitor (Solarbio, Beijing, China), and 1× protease inhibitor cocktail (Solarbio)). For MeRIP, total RNA was isolated using TRIzol. Nuclei pullets or RNA were sheared via sonication at high power for 30 cycles of 5 s on and 30 s off. After that, the supernatant was incubated with 1 μg of antibodies against m6A (Cat. No. ab284130, Abcam) or G3BP1 (Cat. No. 13057-2-AP, Proteintech, Wuhan, Hubei, China) overnight and 40 μl of protein A/G beads (Thermo Fisher Scientific) for 2 h with gentle rotation at 4°C, while the remaining 10% was harvested as input. The pellet beads were centrifuged at 3,000 rpm for 3 min and washed three times. Both the input and pellet beads were purified by TRIzol, and reverse transcription was conducted using a HiScript III 1st Strand cDNA Synthesis Kit (Roche).
description
neonatal
developmental stage
Negative control
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Anas platyrhynchos
organism
Anas platyrhynchos
organism
bursa of Fabricius
organism part
Anas platyrhynchos
scientific_name
male
sex
muscovy ducklings
strain
G3BP1_NC_IP_2
title