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US 2006/0223075 A1Published application
Unique sequence hybridization probes (USP)
AssigneeExagen Diagnostics, Inc.
InventorsLisa Davis, Lei Tang
Life of this patent
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Abstract
The present invention provides unique sequence probes, methods for generating them, and methods for using unique sequence probes in hybridization assays.
Claims13
A method for preparing unique sequence probes comprising:
- (a) identifying unique sequence elements contained in a nucleic acid probe, wherein the nucleic acid probe comprises a plurality of unique sequence elements and a plurality of repetitive sequence elements, and wherein the unique sequence elements in the nucleic acid probe selectively bind to a nucleic acid target of interest;
- (b) amplifying unique sequence elements in the nucleic acid probe to generate amplified unique sequence elements, wherein the amplifying comprises
- (i) contacting the nucleic acid probe with primer sets for a plurality of unique sequence elements under conditions to promote hybridization of the primer sets to the nucleic acid probe, wherein each primer in the plurality of primer sets has an annealing temperature of between 52° C. and 65° C., and wherein the annealing temperature for all of the primers in the plurality of primer sets are within 3° C. of each other;
- and
- (ii) amplifying unique sequence elements defined by the primer sets in by polymerase chain reaction under conditions that comprise annealing the plurality of primer sets to the nucleic acid probe at a temperature between 51° C. and 65° C.;
- and
- (c) pooling the amplified unique sequence elements to produce the unique sequence probe.
4 more elements
A unique sequence probe selected from the group consisting of:
- (a) at least 50 nucleic acids selected from the group consisting of SEQ ID NOS:
- 893-986;
- (b) at least 50 nucleic acids selected from the group consisting of SEQ ID NOS:
- 987-1082;
- (c) at least 50 nucleic acids selected from the group consisting of SEQ ID NOS:
- 1083-1154;
- (d) at least 50 nucleic acids selected from the group consisting of SEQ ID NOS:
- 1155-1242;
- and
- (e) at least 50 nucleic acids selected from the group consisting of SEQ ID NOS:
- 1243-1338.
8 more elements
Isolated polynucleotide primer sets selected from the group consisting of
- (a) at least 1 isolated polynucleotide primer set selected from polynucleotide primer sets 1-94 listed in FIG. 3;
- (b) at least 1 isolated polynucleotide primer set selected from polynucleotide primer sets 1-96 listed in FIG. 4;
- (c) at least 1 isolated polynucleotide primer set selected from polynucleotide primer sets 1-72 listed in FIG. 5;
- (d) at least 1 isolated polynucleotide primer set selected from polynucleotide primer sets 1-88 listed in FIG. 6;
- and
- (e) at least 1 isolated polynucleotide primer set selected from polynucleotide primer sets 1-96 listed in FIG. 7.
3 more elements
Dependencies are inferred from the claim text.