Model organism or animal
BioSampleModel
2023-06-14T23:22:37Z
ENA-FIRST-PUBLIC
2023-06-14T23:22:37Z
ENA-LAST-UPDATE
SAMN26583198
External Id
pda|friendsforvijay
INSDC center name
2023-06-14T23:22:37Z
INSDC first public
2023-06-14T23:22:37Z
INSDC last update
public
INSDC status
SRS12250162
SRA accession
B10.RIII
Submitter Id
13 weeks
age
Rachel Caspi, National Institutes of Health, National Eye Institute, Laboratory of Immunology, Immunoregulation Section, 9000 Rockville Pike, Bldg. 10, Room 10S243, Bethesda, Maryland 20892, USA
biomaterial_provider
7/27/21
birth_date
The Jackson Laboratory
birth_location
NCBI
broker name
Splenocytes
cell_type
Yingyos Jittayasothorn
collected_by
2021-10-26
collection_date
house mouse
common name
The female B10.RIII mouse was purchased from The Jackson Laboratory (ME, USA, strain #: 000457). The animal was maintained under specific-pathogen free (SPF) conditions and euthanized at 13 weeks of age by Carbon dioxide (CO2) inhalation. Splenocytes were isolated using 1X PBS (Gibco), ACK lysing Buffer (Quality Biological), FCS serum (HyClone), RPMI medium (HyClone), DNA Stabilizer (Bionano Genomics), and Cell Buffer (Bionano Genomics). Single cell suspension of 6.8 million Splenocytes were used for high molecular weight genomic DNA (HMW gDNA) extraction, using MagAttract HMW DNA Kit from Qiagen (Cat. No. 67563) for PacBio. Ultra-high molecular weight (UHMW) gDNA was extracted from mouse splenocytes with the Bionano Prep SP Blood and Cell DNA Isolation Kit (Bionano Genomics, 80030) as described in the Bionano Prep SP Frozen Cell Pellet DNA Isolation Protocol (Bionano Genomics). Brifely, 5 million frozen pelleted cells were thawed in a 37oC bath and resuspended in DNA Stabilizing Buffer (Bionano Genomics). After that, the cells were lysed in the presence of detergents, proteinase K, and RNase A and the UHMW gDNA was bound to a silica Nanobind Disk (Bionano Genomics), washed, and eluted. The extracted gDNA was equilibrated for 3 days at room temperature to homogenize and quantified with Qubit dsDNA BR assay kit (Thermo Fisher Scientific, Q32850) and Femto Pulse (Agilent).
description
Healthy
health_state
Mus musculus
organism
Freshly isolated splenocytes
sample_type
Mus musculus
scientific_name
female
sex
B10.RIII
strain
Spleen
tissue
Genomic DNA from B10.RIII mouse splenocytes
title
No
treatment