Sample 10
Gallus gallus
Protocols: Animals were sacrificed at the age of 48 weeks by rapid decapitation. Brains were dissected from the skull and snap frozen in liquid nitrogen within ten minutes, DNA was extracted from hypothalamus using an Allprep RNA/DNA kit (Qiagen), following the manufacturer's instructions. In short, approximately 20-30 mg tissue was homogenized with 600 µl Buffer RTL Plus using FastPrep® -24 (MP Biomedicals). DNA was separated from the RNA using an AllPrep DNA spin column, and columns were kept on ice whilst RNA was purified for gene expression analysis [38]. 350 µl Buffer AW1 was added to the spin column, which was then centrifuged for 15 s at 17000 g. The supernatant was discarded and a mixture of 20 µl proteinase K (20mg/ml) and 60 µl AW1 was added to each sample, and the samples were then incubated for 5 min at room temperature. The samples were then cleaned with 350 µl AW1, centrifuged at 15 s at full speed 17000 g, followed by centrifugation with 500 µl AW2 for 2min at full speed. Columns were dried via an additional centrifugation at 17000 g for 1 min, then placed in 1.5 ml microcentrifuge tubes. For elution of DNA, 50 µl Buffer EB was added to the spin column, followed by a 10 min incubation at room temperature, then centrifugation for 1 min at 8000 g. DNA concentration and purity was measured using a NanoDrop® ND-2000c (ThermoFisher Scientific). DNA methylation analysis was performed by enrichment of the methylated fraction of the genome by immunoprecipitation with an anti-methyl-cytosine antibody (MeDIP). DNA was fragmented using sonication, and captured with agarose beads.
ENA first public
2018-05-29
ENA last update
2018-01-31
External Id
SAMEA104556779
INSDC center alias
Linkoping University
INSDC center name
Linkoping University
INSDC first public
2018-05-29T17:01:38Z
INSDC last update
2018-01-31T09:37:44Z
INSDC status
public
Submitter Id
E-MTAB-6407:Sample 10
age
336
broker name
ArrayExpress
common name
chicken
organism part
hypothalamus
phenotype
low fear
sample name
E-MTAB-6407:Sample 10
sex
female
strain
Red Junglefowl