Methods for solid-phase amplification of DNA template (SPADT) using multiarrays
First Claim
Patent Images
1. A method of detecting the presence or absence of a specific nucleic acid in a sample comprising DNA, said method comprising performing an amplification reaction, wherein a solid support on which a 5′
- and a 3′
primer are irreversibly bound and said DNA is absorbed such that it is reversibly bound, is incubated under amplification conditions, and determining whether said specific nucleic acid is amplified, wherein when said specific nucleic acid is amplified, said specific nucleic acid is present in said sample and when said specific nucleic acid is not amplified, said specific nucleic acid is not present in said sample.
1 Assignment
0 Petitions
Accused Products
Abstract
The present invention relates to a novel method of detecting specific nucleic acids in a biological sample using solid-phase amplification of DNA template (SPADT) using multiarrays.
-
Citations
19 Claims
-
1. A method of detecting the presence or absence of a specific nucleic acid in a sample comprising DNA, said method comprising performing an amplification reaction, wherein a solid support on which a 5′
- and a 3′
primer are irreversibly bound and said DNA is absorbed such that it is reversibly bound, is incubated under amplification conditions, and determining whether said specific nucleic acid is amplified, wherein when said specific nucleic acid is amplified, said specific nucleic acid is present in said sample and when said specific nucleic acid is not amplified, said specific nucleic acid is not present in said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
- and a 3′
-
15. A method of determining the presence or absence of expression of a specific nucleic acid in a cell, said method comprising reverse transcribing RNA obtained from said cell to produce DNA, performing an amplification reaction, wherein a solid support on which a 5′
- and a 3′
primer are irreversibly bound and said DNA is adsorbed such that it is reversibly bound is incubated under amplification conditions, and determining whether said DNA is amplified, wherein when said DNA is amplified, said specific nucleic acid is expressed in said cell and when said DNA is not amplified, said specific nucleic acid is not expressed in said cell.
- and a 3′
-
16. A method of determining the presence or absence of a specific RNA molecule in a sample, said method comprising reverse transcribing said RNA to DNA, performing an amplification reaction wherein a solid support on which a 5′
- and a 3′
primer are irreversibly bound and said DNA is adsorbed such that it is reversibly bound is incubated under amplification conditions, and determining whether said DNA is amplified, wherein when said DNA is amplified, said specific RNA is present in said sample, and when said DNA is not amplified, said specific RNA is not present in said sample. - View Dependent Claims (17)
- and a 3′
-
18. A method of designing a primer which amplifies a specific nucleic acid provided said specific nucleic acid is present in a sample comprising DNA, said method comprising
(a) synthesizing at least one 5′ - and one 3′
primer homologous to said specific nucleic acid;
(b) performing an amplification reaction wherein a solid support on which said primer is irreversibly bound and said DNA is absorbed such that it is reversibly bound is incubated under amplification conditions to generate an amplified nucleic acid;
(c) determining the specificity of said primer by comparing the ability of said primer to produce a detectable amount of said amplified nucleic acid in said amplification reaction in the presence or absence of said specific nucleic acid, wherein a detectable amount of amplified nucleic acid in said amplification reaction in the presence of said specific nucleic acid, compared with the absence of a detectable amount of said amplified nucleic acid in the absence of said specific nucleic acid, is an indicator that said primer is specific for ampliyfing said specific nucleic acid;
(d) comparing the characteristics of said primers of step (c) which are specific for said specific nucleic acid to identify common features shared among the most specific of said primers; and
(e) synthesizing an additional primer incorporating said features identified in step (d), thereby designing a primer which amplifies a nucleic acid provided said specific nucleic acid is present in a sample comprising DNA. - View Dependent Claims (19)
- and one 3′
Specification