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US 5,512,439 AGranted
Oligonucleotide-linked magnetic particles and uses thereof
AssigneeDynal AS
InventorsErik Hornes, Lars Korsnes
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Abstract
Monodisperse, superparamagnetic particles carrying a plurality of molecules of an oligonucleotide are disclosed and may be used inter alia for sequencing single-stranded nucleic acids. The oligonucleotide may be covalently attached or affinity bonded to the particles either by their 3' or 5' termini. The particles have a specific gravity in the range 1.1-1.8 and are 1-10 microns in diameter. A kit for the isolation or processing of target nucleic acid is also disclosed.
Claims20
A plurality of monodisperse, superparamagnetic particles, wherein:
- each particle comprises (i) superparamagnetic iron oxide dispersed within a polymer particle, (ii) a coating which reduces non specific binding, and (iii) a functional group carried by said coating for bonding a nucleic acid
- the particles are monodisperse and have a diameter standard deviation of less than 5%, and
- particles of said plurality carry a plurality of molecules of an oligonucleotide.
A method of sequencing single stranded nucleic acids which comprises the steps of:
- (a) preparing a plurality of superparamagnetic monodisperse particles wherein each particle comprises (i) superparamagnetic iron oxide dispersed within a polymer particle and (ii) a coating which reduces non-specific binding;
- and which carries a functional group for bonding a nucleic acid and the particles being monodisperse and having a diameter standard deviation of less that 5%, and particles of said plurality carrying a plurality of moleceles of an oligonucleotide to be sequenced;
- (b) either
- (i) dividing the particles into four aliquots and adding to each aliquot a polymerase, mixed nucleoside triphosphates, a single dideoxynucleoside triphosphate, the latter being different for each aliquot and, a primer, if required, wherein at least one of the primer or nucleoside or dideoxynucleoside is labelled;
- or
- (ii) adding to all the particles a polymerase, mixed nucleoside triphosphates, four different dideoxy nucleoside triphosphates each carrying a different label and, a primer, whereby, a series of labelled DNA strands are synthesized each having different chain lengths and ending with a particular dideoxy base,
- (c) liberating the labelled DNA strands and size fractionating these;
- and
- (d) determining the sequence of the nucleic acids.
6 more elements
Dependencies are inferred from the claim text.