Rapid purification of nuclei from animal and plant tissues and cultured cells.
Nuclei are isolated by buoyant density banding in a discontinuous iodixanol gradient, under isoosmotic conditions. The low viscosity of the gradient allows the purification to be carried out at 10,000g in only 20 min. The method avoids possible damage to nucleoprotein complexes caused by hyperosmotic sucrose gradients. Although developed for mammalian liver the method can be applied (with or without minor modifications) to any tissue or cell type.
Graham
John M
JM
School of Biomolecular Sciences, Liverpool John Moores University. john@jgrescon.fsbusiness.co.uk
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002467:Cell Nucleus; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient / Q000379:methods*; D008099:Liver / Q000648:ultrastructure; D010944:Plants / Q000648:ultrastructure; D014283:Triiodobenzoic Acids / Q000737:chemistry
D014283:Triiodobenzoic Acids; C044834:iodixanol