2025-04-01
ENA first public
ERC000011
ENA-CHECKLIST
Department of rheumatology Tokyo Medical and Dental University (TMDU)
INSDC center name
public
INSDC status
ERS22446151
SRA accession
E-MTAB-14621:IGR_Lung_Early3
Submitter Id
19
week
age
ArrayExpress
broker name
not collected
collection date
house mouse
common name
Protocols: Mouse-adapted SARS-CoV-2 strain was established in the previous study. Mice were anesthetized and then inoculated viral solution intranasally (i.n.). Viral solutions for inoculation were diluted with PBS and 30 µl of solution contained twice of the 50% lethal dose (2LD50) determined in 24-week-old BALB/c mice. At three post infection, mice were anesthetized by intraperitoneal injection with a mixture of 0.75 mg/kg of medetomidine, 4.0 mg/kg of midazolam, and 5.0 mg/kg of butorphanol. Lung samples were dissected from mice and immediately immersed in RNA later (Invitrogen) stabilization reagent and stored at -80℃ for 24 hr. Samples were transferred to a new tube containing 1-mm glass beads and 700μL Trizol (Invitrogen). Tissues were subsequently homogenized using a MINI BEAD-BEATER (Biospec). Total RNA was extracted and purified using a RNeasy mini kit (QIAGEN, Tokyo, Japan) and quantified using a spectrophotometer (NanoDrop one, Thermo Fisher Scientific).  Total RNA was extracted from the lung as described above and checked for quality using RNA integrity scores were typically 7.0 and greater with an Agilent 2100 bioanalyzer (Agilent, Santa Clara, USA). For the quality control, mRNA had to meet the requirement of 28S/18S > 1, while miRNA had to meet the requirement of 28S/18S > 1.5. According to the manufacturer's protocol, the RNA was fragmented into small pieces. Afterward, the fragmented RNA was reverse-transcribed into cDNA and amplified with polymerase chain reaction (PCR) to create a cDNA library. Quality control and quantification of the libraries were performed with the Agilent 2100 bioanalyzer and real-time quantitative PCR (qPCR) (TaqMan Probe).
description
adult
developmental stage
high fat diet
diet
Mouse-adapted SARS-CoV-2, 3 days post infection
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
lung
organism part
Mus musculus
scientific_name
male
sex
C57BL/6
strain
IGR_Lung_Early3
title