Solid phase for capture of nucleic acids

摘要

A method of: providing a solid surface having a dendrimer molecule bound thereto and a single-stranded probe nucleic acid immobilized to the dendrimer; contacting the solid surface with a sample suspected or known to contain a double-stranded complimentary target nucleic acid; denaturing the target nucleic acids at thermal conditions and in a salt concentration sufficient to denature the target nucleic acids to produce denatured nucleic acids; and cooling the sample to allow hybridization of the denatured nucleic acids to the probe nucleic acids. An article having: one or more paramagnetic microbeads; a dendrimer molecule bound to the beads; and a probe nucleic acid immobilized to the dendrimer.

主权项

1. A method comprising:
providing a solid surface having a dendrimer molecule covalently bound thereto and a single-stranded probe nucleic acid immobilized to the dendrimer;
wherein the dendrimer is N3P3(O—C6H4—X)6 or: wherein n is a positive integer;wherein N3P3 is hexavalent cyclotriphosphazene;wherein the repeating unit in parentheses indicates that phosphorous atom on the right is bound to two repeat units;wherein each X is independently selected from —CHO and —CH2—NH—;wherein each —CH2—NH— group is directly or indirectly bound to the solid surface or the probe nucleic acid; andwherein there is at least one —CH2—NH— group directly or indirectly bound to the solid surface and at least one —CH2—NH— group directly or indirectly bound to the probe nucleic acid; contacting the solid surface with a sample suspected or known to contain a double-stranded target nucleic acid;
wherein the solid surface comprises a glass slide, a silicon wafer, a structured silicon wafer, or a plurality of paramagnetic, glass, or silica beads;wherein a strand of the target nucleic acid is complementary to the probe nucleic acid; denaturing the target nucleic acids while the sample is in contact with the solid surface at thermal conditions and in a salt concentration sufficient to denature the target nucleic acids to produce denatured nucleic acids;
wherein the-dendrimer molecule remains covalently bound to the solid surface at the thermal conditions and the salt concentration; and cooling the sample to allow hybridization of the denatured nucleic acids to the probe nucleic acids.