cfa3
mixed sample
Protocols: doi:10.1016/j.ymeth.2009.03.001; Liver material was crosslinked in 1% formaldehyde for 20 minutes followed by a 10 minute incubation in 500 mM glycine buffer to neutralize the formaldehyde. Samples were rinsed with PBS and frozen until use. doi:10.1016/j.ymeth.2009.03.001; Crosslinked animal livers were dounce homogenized into a single cell suspension and rinsed with ice cold PBS. Each pellet of cross-linked material was resuspended in 10 ml of LB1 (50 mM Hepes-KOH, pH 7.5; 140 mM NaCl; 1mM EDTA; 10% Glycerol; 0.5% NP-40 or Igepal CA-630; 0.25% Triton X-100) and pelleted again by centrifugation at 2,000 x rcf for 4 minutes at 4 C. Pellets were then rinsed in 10 ml of LB2 (10 mM Tris-HCL, pH8.0; 200 mM NaCl; 1 mM EDTA; 0.5 mM EGTA) and pelleted again by centrifugation at 2,000 x rcf for 4 minutes at 4 C. The nuclei preparation was resuspended in 3 ml LB3 (10 mM Tris-HCl, pH 8; 100 mM NaCl; 1 mM EDTA; 0.5 mM EGTA; 0.1% Na-Deoxycholate; 0.5% N-lauroylsarcosine). Chromatin in the nuclei fraction was fragmented to an average length of 300 bp by sonication. After sonication 300 ul of 10% Triton X-100 was added to the 3 ml of sonicated lysate and debris was removed by centrifugation at 20,000 x rcf for 10 minutes at 4 C.
DevelopmentalStage
adult
ENA-FIRST-PUBLIC
2013-03-06
ENA-LAST-UPDATE
2018-03-08
External Id
SAMEA1978323
INSDC center alias
CRI
INSDC center name
Cancer Research UK Cambridge Research Institute
INSDC first public
2013-03-06T17:02:06Z
INSDC last update
2018-03-08T15:33:37Z
INSDC status
suppressed
Individual
cfa3
OrganismPart
liver
Sex
male
Submitter Id
E-MTAB-890:cfa3
broker name
ArrayExpress
common name
dog
sample name
E-MTAB-890:cfa3
component_organism
Canis lupus familiaris
component_organism
Canis lupus familiaris