Rapid isolation of polynucleotides

Details Rapid isolation of polynucleotides Rapid isolation of polynucleotides

1995-05-25

1997-04-29

Kight, John

Organic compounds -- part of the class 532-570 series

Organic compounds

Carbohydrates or derivatives

536 254, 536 2541, C07H 2104

Patent

active

056250550

ABSTRACT:
Polynucleotides (particularly DNA) formed from a lysate of cellular material in a holding tube are separated from proteins in a two-phase solvent system formed by a mild oxidising perchlorate and an organic solvent such as chloroform. A powder of polymerised silica gel particles containing free aldehyde (CHO) or ketone (CO) groups which react with the proteins is added to the uppermost layer which is the layer containing the polynucleotides of interest and is allowed to descend through that layer thereby cleansing it of protein contaminants. The protein reacted silica gel particles lodge at the interface between the upper polynucleotide containing layer and the lower organic solvent layer which contains the bulk of the proteins and forms a solid disc adherent to the walls of the holding tube. The uppermost layer can thereafter be decanted for precipitation of the polynucleotides therein without contamination from the protein containing solvent layer. The powder of polymerised silica gel particles is preferably prepared by cross-linking silica gel particles of particle size 250-400 mesh with 3-aminopropyltriethosilane followed by reaction with gluteraldehyde.

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Vandenberg et al., "Protein Immobilization to 3-Aminopropyl Triethoxysilance/Glutaraldehyde Surfaces: Characterization by Detergent Washing," J. Colloid Interface Sci., 143(2), 327-335 (1991); Chem. Abstr., 114(23), p. 457, Abstr. No. 225132s (1991); only Abstract provided.
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