Bulk isolation in nonaqueous media of nuclei from lyophilized cells.
Intact lyophilized nuclei are obtainable from a variety of tissues, either in situ or in culture, by freezing at -156 degrees C, drying at -25 degrees C, and mechanical disassociation in glycerol at 2 degrees C. Centrifugal separation of nuclei is accomplished in an 85 : 15 by volume mixture of glycerol and 3-chloro-1,2 propanediol at 2 degrees C. The method gives homogeneous nuclear preparations in high yield with preservation of labile and water-soluble constituents.
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D006528:Carcinoma, Hepatocellular; D002467:Cell Nucleus / Q000032:analysis*; D002645:Chickens; D004247:DNA / Q000032:analysis; D004912:Erythrocytes / Q000166:cytology; D005612:Freeze Drying; D005909:Glioblastoma; D005947:Glucose / Q000032:analysis; D005990:Glycerol; D006019:Glycolysis; D006600:Hexosephosphates / Q000032:analysis; D007773:Lactates / Q000032:analysis; D008099:Liver / Q000166:cytology; D008113:Liver Neoplasms; D008722:Methods; D051379:Mice; D008854:Microscopy, Electron; D011409:Propylene Glycols; D012313:RNA / Q000032:analysis
D006600:Hexosephosphates; D007773:Lactates; D011409:Propylene Glycols; D012313:RNA; D000255:Adenosine Triphosphate; D004247:DNA; D005947:Glucose; D005990:Glycerol