Fractionation of chromosomes. II. Use of hydrophobic affinity partition in aqueous two-phase system.
A simple method for separation of large quantities of isolated metaphase chromosomes in Single-Tube Partition (STP), using hydrophobic ligand in an aqueous two-phase system is presented. The two-phase system is composed of an aqueous solution of Dextran 500 and poly(ethylene) glycol 6000 (PEG). The concentration of chromosomes to be separated has no influence on the distribution behaviour in the partition system and up to 10(7) chromosomes can be used in a phase system as small as 3-5 g (5 ml tube). Different groups of chromosomes differ in their distribution in the two phases and the introduction of PEG with covalently attached hydrophobic ligand provides a means of controlling the distribution of chromosomes. A combination of positively charged trimethylaminomethane PEG (TMA-PEG) together with palmitat PEG (P-PEG) gives a fairly good condition for separating chromosomes on the basis of their net surface charge differences.
D002458:Cell Fractionation / Q000379:methods*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002877:Chromosomes, Human / Q000648:ultrastructure; D003911:Dextrans; D006367:HeLa Cells; D006801:Humans; D008677:Metaphase; D011092:Polyethylene Glycols; D000644:Quaternary Ammonium Compounds; D013499:Surface Properties
D003911:Dextrans; D000644:Quaternary Ammonium Compounds; D011092:Polyethylene Glycols; C020470:polyethyleneglycol palmitate; C027917:tetramethylammonium