title: lean ob_3
organism: Mus musculus
description: 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).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-05-15T15:00:26Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-05-15T15:00:26Z
attributes.tag: External Id
attributes.value: SAMEA112651711
attributes.tag: INSDC center name
attributes.value: Department of rheumatology Tokyo Medical and Dental University (TMDU)
attributes.tag: INSDC first public
attributes.value: 2023-05-15T15:00:26Z
attributes.tag: INSDC last update
attributes.value: 2023-05-15T15:00:26Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12623:lean ob_3
attributes.tag: age
attributes.value: 16
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: leptin deficient
attributes.tag: infect
attributes.value: Severe acute respiratory syndrome coronavirus 2
attributes.tag: organism part
attributes.value: lung
attributes.tag: phenotype
attributes.value: lean
attributes.tag: sample name
attributes.value: E-MTAB-12623:lean ob_3
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57BL/6JHamSlc-ob/ob