Labelling of oligonucleotides

Claim 2

1 . A method of making an oligonucleotide which comprises hybridizing (a) a shorter fragment of the desired oligonucleotide with (b) a nucleic acid fragment longer than (a) and complementary to the desired oligonucleotide, one of (a) and (b) carrying an organic radical which differentiates its physical properties relative to the other of (a) and - (b), contacting the hybridized material with an enzyme and nucleoside triphosphates whereby the shorter fragment grows in one direction until it is substantially coterminus with the complementary nucleic acid fragment, denaturing the hybridized material and separating lengthened (a) from (b).

Claim 3

The method according to claim 1, wherein the organic radical is a reactive site and separation is effected by contact with a medium which reacts with the reactive site.

Claim 8

A hybridized material comprising (b) a nucleic acid fragment and (a) a shorter oligonucleotide hybridized thereto, one of (a) and (b) carrying an organic radical which differentiates its physical or chemical properties relative to the other of (a) and (b).

Claim 9

A hybridized material comprising (b) a nucleic acid fragment and (a) a shorter oligonucleotide hybridized thereto, one of (a) and (b) carrying an organic radical which differentiates its physical or chemical properties relative to the other of (a) and (b).

Claim 10

A hybridized material comprising two short fragments of nucleic acid and one of the following structures: wherein the X:Y is T:A for the sickle cell probe and A:T for the corresponding normal globin gene probe, b) hemoglobin C wherein the X:Y is A:T for the Hb C probe and G:C for the corresponding normal globin gene probe, c) beta° (B 39 ) thalassemia wherein the X is A for the beta° probe and G for the corresponding normal globin gene probe, d) (i) beta + thalassemias wherein X, Y and Z are G, G and T for the normal globin gene probe, A, G and T for the IVS-1 thalassemia probe, G, C and T for the IVS-5 thalassemia probe, and G, G and C for the IVS-6 thalassemia probe; N is the appropriate complementary nucleotide, (ii) beta + thalassemias wherein X:Y is A:T for the IVS-110 thalassemia probe and G:C for the corresponding normal globin gene probe, e) homozygous alpha-thalassemia f) alpha, protease inhibitor (antitrypsin) deficiency wherein X:Y is A:T for the deficiency (Z) probe and G:C for the corresponding normal globin gene (M) probe.

Claim 11

A hybridized material comprising two short fragments of nucleic acid and one of the following structures: wherein the X:Y is T:A for the sickle cell probe and A:T for the corresponding normal globin gene probe, b) hemoglobin C wherein the X:Y is A:T for the Hb C probe and G:C for the corresponding normal globin gene probe, c) beta° (B 39 ) thalassemia wherein the X is A for the beta° probe and G for the corresponding normal globin gene probe, d) (i) beta + thalassemias wherein X, Y and Z are G, G and T for the normal globin gene probe, A, G and T for the IVS-1 thalassemia probe, G, C and T for the IVS-5 thalassemia probe, and G, G and C for the IVS-6 thalassemia probe; N is the appropriate complementary nucleotide, (ii) beta + thalassemias wherein X:Y is A:T for the IVS-110 thalassemia probe and G:C for the corresponding normal globin gene probe, e) homozygous alpha-thalassemia f) alpha, protease inhibitor (antitrypsin) deficiency wherein X:Y is A:T for the deficiency (Z) probe and G:C for the corresponding normal globin gene (M) probe.

Claim 12

An assay system for detection of a nucleic acid in a biological sample containing an oligonucleotide which is prepared by a method which comprises hybridizing (a) a shorter fragment of the desired oligonucleotide with (b) a nucleic acid fragment longer than (a) and complementary to the desired oligonucleotide, one of (a) and (b) carrying an organic radical which differentiates its physical properties relative to the other of (a) and (b), contacting the hybridized material with an enzyme and nucleoside triphosphates whereby the shorter fragment grows in one direction until it is substantially coterminus with the complementary nucleic acid fragment, denaturing the hybridized material and spearating lengthened (a) form (b).

Claim 13

An assay system for detection of a nucleic acid in a biological sample containing an oligonucleotide which is prepared by a method which comprises hybridizing (a) a shorter fragment of the desired oligonucleotide with (b) a nucleic acid fragment longer than (a) and complementary to the desired oligonucleotide, one of (a) and (b) carrying an organic radical which differentiates its physical properties relative to the other of (a) and (b), contacting the hybridized material with an enzyme and nucleoside triphosphates whereby the shorter fragment grows in one direction until it is substantially coterminus with the complementary nucleic acid fragment, denaturing the hybridized material and spearating lengthened (a) form (b).

Claim 14

Use of an oligonucleotide in a nucleic acid hybridization assay which is prepared by a method which comprises hybridizing (a) a shorter fragment of the desired oligonucleotide with (b) a nucleic acid fragment longer than (a) and complementary to the desired oligonucleotide, one of (a) and (b) carrying an organic radical which differentiates its physical properties relative to the other of (a) and - (b), contacting the hybridized material with an enzyme and nucleoside triphosphates whereby the shorter fragment grows in one direction until it is substantially coterminus with the complementary nucleic acid fragment, denaturing the hybridized material and separating lenghtened (a) from (b).

Claim 15

Use of an oligonucleotide in a nucleic acid hybridization assay which is prepared by a method which comprises hybridizing (a) a shorter fragment of the desired oligonucleotide with (b) a nucleic acid fragment longer than (a) and complementary to the desired oligonucleotide, one of (a) and (b) carrying an organic radical which differentiates its physical properties relative to the other of (a) and - (b), contacting the hybridized material with an enzyme and nucleoside triphosphates whereby the shorter fragment grows in one direction until it is substantially coterminus with the complementary nucleic acid fragment, denaturing the hybridized material and separating lenghtened (a) from (b).