2024-10-31
ENA first public
ERC000011
ENA-CHECKLIST
Vanderbilt University
INSDC center name
public
INSDC status
ERS18638546
SRA accession
E-MTAB-13982:KL2
Submitter Id
ArrayExpress
broker name
not collected
collection date
house mouse
common name
Protocols: Embryos were generated by crossing a single housed 8 week old heteozygous male mouse with a heterozygous 6-8 week old female mouse. Female mice with vaginal plugs were defined as being E0.5. At the E12.5 time point, embryonic tissues were micro-dissected in cold phosphate buffered saline (PBS) and embryos staged according to Theiler staging criteria. Sox17∆50  (Sox17em3Mgn) mutant allele was generated using CRISPR/Cas9 mutagenesis by pronuclear injection into  fertilized zygotes from the mating of CD-1 mice. Founder animals were backcrossed to wild type CD-1 mice for three generations before interbreeding to produce homozygous mutant mice.    Mice were fed a regular chow  (4.5% fat content, PicoLab, 5L0D) diet. Total RNA was extracted from isolated mouse pancreatic islets using the Maxwell simplyRNA kit (Promega). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. Final library was complete after end repair, A-tailing, adapter ligation, size selection, amplification, and purification.The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.
description
E12.5
developmental stage
Sox17D50/D50
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
liver
organism part
Mus musculus
scientific_name
Undetermined
sex
CD1
strain
KL2
title