Methods for analysis of gene expression
First Claim
Patent Images
1. A method for analyzing gene expression comprising:
- a) obtaining a plurality of target sequences, wherein the plurality of target sequences comprises cDNA;
b) multiplex amplifying said plurality of target sequences, wherein multiplex amplifying comprises combining the plurality of target sequences, a plurality of target-specific primers, and one or more universal primers, thereby producing a plurality of amplification products;
c) separating one or more members of the plurality of amplification products;
d) detecting one or more members of the plurality of amplification products, thereby generating a set of gene expression data;
e) storing the set of gene expression data in a database; and
f) performing a comparative analysis on the set of gene expression data, thereby analyzing the gene expression.
1 Assignment
0 Petitions
Accused Products
Abstract
This invention provides methods, compositions and kits for gene expression analysis and gene expression profiling. The methods of the invention are highly sensitive; have a wide dynamic range; are rapid and inexpensive; have a high throughput; and allow the simultaneous differential analysis of a defined set of genes. The methods, compositions and kits of the invention also provide tools for gene expression data collection and relational data analysis.
-
Citations
107 Claims
-
1. A method for analyzing gene expression comprising:
-
a) obtaining a plurality of target sequences, wherein the plurality of target sequences comprises cDNA;
b) multiplex amplifying said plurality of target sequences, wherein multiplex amplifying comprises combining the plurality of target sequences, a plurality of target-specific primers, and one or more universal primers, thereby producing a plurality of amplification products;
c) separating one or more members of the plurality of amplification products;
d) detecting one or more members of the plurality of amplification products, thereby generating a set of gene expression data;
e) storing the set of gene expression data in a database; and
f) performing a comparative analysis on the set of gene expression data, thereby analyzing the gene expression. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 87)
-
-
59. A method for analyzing gene expression comprising:
-
a) obtaining cDNA from a plurality of samples for a plurality of target sequences;
b) performing a plurality of multiplexed amplifications of the target sequences, thereby producing a plurality of multiplexed amplification products;
c) pooling the plurality of multiplexed amplification products;
d) separating the plurality of multiplexed amplification products;
e) detecting the plurality of multiplexed amplification products, thereby generating a set of gene expression data;
f) storing the set of gene expression data in a database; and
g) performing a comparative analysis of the set of gene expression data. - View Dependent Claims (60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 88)
-
-
75. A method for analyzing gene expression comprising:
-
a) obtaining cDNA from multiple samples;
b) amplifying a plurality of target sequences from the cDNA, thereby producing a multiplex set of amplification products;
c) separating and detecting the amplification products using a high throughput platform, wherein detecting generates a set of gene expression data; and
d) storing the set of gene expression data in a database; and
e) performing a comparative analysis of the set of gene expression data. - View Dependent Claims (76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 89, 105, 106, 107)
-
-
90. A system for analyzing gene expression, the system comprising:
-
a) an amplification module for producing a plurality of amplification products from a pool of target sequences, the amplification module comprising at least one pair of universal primers and at least one pair of target-specific primers;
b) a detection module for detecting one or more members of the plurality of amplification products, wherein the detection module detects a presence, absence, or quantity of the one or more members, and generates a set of gene expression data comprising a plurality of data points; and
c) an analyzing module in operational communication with the detection module, the analyzing module comprising a computer or computer-readable medium comprising one or more logical instructions which organize the plurality of data points into a database and one or more logical instructions which analyze the plurality of data points. - View Dependent Claims (91, 92, 93, 94, 95, 96, 97, 98, 99, 100)
-
-
101. A composition for preparing a plurality of amplification products from a plurality of mRNA target sequences, the composition comprising:
-
one or more pairs of universal primers; and
one or more pairs of target-specific primers, wherein the target-specific primers comprise one or more regions complementary to the one or more pairs of universal primers and one or more regions complementary to one or more target mRNA sequences. - View Dependent Claims (102, 103)
-
-
104. A kit for obtaining a multiplex set of amplification products of target genes and references-genes, the kit comprising:
-
a) at least one pair of universal primers;
b) at least one pair of target-specific primers;
c) at least one pair of reference gene-specific primers; and
d) one or more amplification reaction enzymes, reagents, or buffers.
-
Specification