2024-06-10
ENA first public
ERC000011
ENA-CHECKLIST
Division of Regulatory Genomics and Cancer Evolution, German Cancer Research Center
INSDC center name
public
INSDC status
ERS20104284
SRA accession
E-MTAB-14096:WTM
Submitter Id
20
week
age
ArrayExpress
broker name
not collected
collection date
house mouse
common name
Protocols: Dissected spleen fragments were transferred to a 40-μm cell strainer (Greiner # 542040) and ground with a syringe plunger. The cell suspension was strained twice. Strainers were washed with additional DMEM (Gibco # 41966029) with 5% FBS and cells were pelleted by centrifugation at 300 x g for 4 minutes at 4°C. Cell pellets were carefully resuspended in 300 uL of ACK lysis buffer (Lonza # BP10-548E) for 1 minute in ice. After incubation cells were washed with 1mL of DMEM with 5% FBS and pelleted. DNA was crosslinked, fragmented, ligated, de-crosslinked and shuffled to get circular templates. Inverse PCR was performed using independent primer sets respectively targeting the 5’ end of the chrX duplication (forward TTCAGTAGGTGTGACAGAGA; reverse ACATAATTATAGCGCTGGCA), 3’ end of the chrX duplication (forward GCATCTGTCTGCTTACTTTG; reverse GGTCTGGATACCTTTCAAGG) and the chrY translocation site identified with the first two pairs (forward TAAAAGGGGAGGTAGACACA; reverse GTGTTCAAACCAGACTCCT). PCR products were purified, library prepped using the Nextera DNA Flex (Illumina) chemistry and pooled.
description
adult
developmental stage
wild type genotype
genotype
not collected
geographic location (country and/or sea)
1416027
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
spleen
organism part
Mus musculus
scientific_name
male
sex
C57BL/6J
strain
WTM
title