US 5,470,707 A · Hydrogen bond labeling and base sequence determination methods for DNA or RNA

Abstract

A hydrogen bond labeling and base sequence determination method for DNA or RNA takes advantage of the complementary nature of the fundamental structure of DNA or RNA bases to bind corresponding chemical species with the base species of a denatured DNA or RNA chain. After providing an aqueous solution containing a nucleic acid, heating the aqueous solution to cleave the nucleic acid; cooling the aqueous solution and adding a base-specific labeling molecule having available hydrogen bonds to the aqueous solution, the base-specific labeling molecules bond to the available bases in a one-to-one fashion via the available hydrogen bonds. The resulting labeled molecule can be microscopically observed to determine the base sequence of the nucleic acid.

Claims17

Dependent claims

A method for labeling a nucleic acid base of a DNA or RNA molecule, comprising the steps of:

preparing an aqueous solution containing a double-stranded DNA or RNA molecule;
heating the aqueous solution to denature the double-stranded DNA or RNA molecule so that the double strand dissociates into single strands having complementary nucleotide sequences;
adding at least one base-specific labeling molecule to the aqueous solution;
cooling the aqueous solution so that the base-specific labeling molecule is hybridized to complementary bases on the single strands to form a contour of each of the single strands defined by said at least one base-specific labeling molecule;
and
further cooling the aqueous solution so that the single strands, having the hybridized base-specific labeling molecule, are independently bound on a substrate;
the contour of each of said independently bound strands, due to the hybridized base-specific labeling molecule, representing a base sequence thereof.

4 more elements

A method for determining a base sequence of a DNA or RNA molecule, comprising the steps of:

preparing first, second and third aqueous solutions containing identical double-stranded DNA or RNA molecules;
heating the first, second, and third aqueous solutions to denature the respective double-stranded DNA or RNA molecules so that each of the double strands dissociates into single strands having complementary nucleotide sequences;
adding at least one first, at least one second, and at least one third base-specific labeling molecule to the first, second, and third aqueous solutions, respectively;
cooling the first, second, and third aqueous solutions so that the respective base-specific labeling molecule added to each aqueous solution is hybridized to complementary bases on the single strands in the respective aqueous solution, to form a contour of each of the single strands defined respectively by said at least one first, at least one second, and at least one third base-specific labeling molecule;
further cooling the first, second, and third aqueous solutions so that each single strand, having the respective hybridized base-specific labeling molecule, is independently bound on a substrate;
the contour of each of said independently bound strands, due to the respective hybridized base-specific labeling molecule, representing a base sequence thereof;
determining the labeled sequence of each of the independently bound single strands from the respective contours thereof;
and
determining the base sequence of the identical double stranded parts of DNA or RNA from the step of determining the labeled sequence of each of the independently bound single strands.

6 more elements

Dependencies are inferred from the claim text.

Hydrogen bond labeling and base sequence determination methods for DNA or RNA

Life of this patent

Hydrogen bond labeling and base sequence determination methods for DNA or RNA

Life of this patent

Abstract

Claims17