Polymers of oligonucleotide probes as the bound ligands for use in reverse dot blots

Claim 1

A method of detecting the presence of nucleic acid sequences, comprising: binding polymers of at least one oligonucleotide probe to a substrate wherein said polymeric probes are comprised of monomeric sequence units that are complementary to a target sequence which exists as a single copy within a region of a nucleotide sequence contained in a DNA test sample wherein said monomeric sequence units are selceted from the group consisting of Sequence I.D. Nos. 1-112; contacting said substrate to which said polymeric probes are bound with amplification products of said test DNA sample containing uncharacterized oligonucleotide sequences, wherein a reporter moiety has been incorporated, such that individual complementary sequences that are contained in the amplified test sample are able to anneal to any monomeric sequence that is contained in any polymeric probe bound to said substrate; washing said substrate to remove unannealed amplified DNA test sample from said substrate; and detecting the presence or absence of reporter moieties retained by said polymeric probes on said substrate.

Claim 2

A method of detecting the presence of nucleic acid sequences, comprising: binding polymers of at least one oligonucleotide probe to a substrate wherein said polymeric probes are comprised of monomeric sequence units that are complementary to a target sequence which exists as a single copy within a region of a nucleotide sequence contained in a DNA test sample wherein said monomeric sequence units are selceted from the group consisting of Sequence I.D. Nos. 1-112; contacting said substrate to which said polymeric probes are bound with amplification products of said test DNA sample containing uncharacterized oligonucleotide sequences, wherein a reporter moiety has been incorporated, such that individual complementary sequences that are contained in the amplified test sample are able to anneal to any monomeric sequence that is contained in any polymeric probe bound to said substrate; washing said substrate to remove unannealed amplified DNA test sample from said substrate; and detecting the presence or absence of reporter moieties retained by said polymeric probes on said substrate.

Claim 5

A method of synthesizing oligonucleotides probes consisting of repeated monomeric sequence units by the steps of: (a) annealing two oligonucleotide primers which are oppositely oriented and which are complementary at their 3' ends and which together either directly or in the form of complementary nucleotides define the sequence of a complete monomeric unit which incorporates the selected oligonucleotide probe sequence as well as a sufficient number of the nucleotides of the adjacent monomeric units such that the primers anneal at their 3' ends and that the 5' ends of the primers are not annealed and function as templates for synthesis of first extension products; (b) treating the primers with a DNA polymerase in the presence of deoxyribonucleotides such that a first extension product of each primer is synthesized which is complementary to each template; (c) separating the complementary first extension products by denaturation to produce single-stranded molecules; (d) annealing complementary strands of said first extension products at their 3' ends in a staggered arrangement such that the 5' ends are not annealed and function as templates for further extension of the previous extension products; (e) treating the annealed strands using a DNA polymerase and deoxyribonucleotides such that double-stranded extension products are synthesized; (f) separating the complementary extension products by denaturization; and (g) repeating steps (d) through (f) using the extension products of step (f) to produce successively longer extension products with each repetition without the addition of any new primer sequences not generated by the initial two oligonucleotide primers.

Claim 6

A method of synthesizing oligonucleotides probes consisting of repeated monomeric sequence units by the steps of: (a) annealing two oligonucleotide primers which are oppositely oriented and which are complementary at their 3' ends and which together either directly or in the form of complementary nucleotides define the sequence of a complete monomeric unit which incorporates the selected oligonucleotide probe sequence as well as a sufficient number of the nucleotides of the adjacent monomeric units such that the primers anneal at their 3' ends and that the 5' ends of the primers are not annealed and function as templates for synthesis of first extension products; (b) treating the primers with a DNA polymerase in the presence of deoxyribonucleotides such that a first extension product of each primer is synthesized which is complementary to each template; (c) separating the complementary first extension products by denaturation to produce single-stranded molecules; (d) annealing complementary strands of said first extension products at their 3' ends in a staggered arrangement such that the 5' ends are not annealed and function as templates for further extension of the previous extension products; (e) treating the annealed strands using a DNA polymerase and deoxyribonucleotides such that double-stranded extension products are synthesized; (f) separating the complementary extension products by denaturization; and (g) repeating steps (d) through (f) using the extension products of step (f) to produce successively longer extension products with each repetition without the addition of any new primer sequences not generated by the initial two oligonucleotide primers.