Methods of assaying differential expression

• US 5,994,076 A
• Filed: 05/21/1997
• Issued: 11/30/1999
• Est. Priority Date: 05/21/1997
• Status: Expired due to Fees

• Current Assignee: Clontech Laboratories Incorporated (Ningbo Junsheng Investment Group Co., Ltd.)
• Original Assignee: Clontech Laboratories Incorporated (Ningbo Junsheng Investment Group Co., Ltd.)
• Inventors: Chenchik, Alex, Jokhadze, George, Bibilashvilli, Robert
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US08/859,998
• Time in Patent Office: 923 Days
• Field of Search: 435/6, 435/91.1, 435/91.2, 536/24.3, 536/24.31, 536/24.33, 536/23.1
• US Class Current: 435/6.16
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof

Abstract

Methods and compositions are provided for analyzing differences in the RNA profiles between a plurality of different physiological samples. In the subject methods, a set of a representational number of distinct gene specific primers is used to generate labeled nucleic acids from each of the different physiological samples. The labeled nucleic acids are then compared to each other and differences in the RNA profiles are determined. The subject methods find use in methods of identifying differential gene expression.

Claims

1. A method of producing a population of labeled nucleic acids, said method comprising;

obtaining a sample of nucleic acids from a physiological source; and

generating a population of labeled nucleic acids from the nucleic acids sample by using a set of primers comprising at least 20 primers selected from SEQ ID NOS;

01-1372;

whereby a population of labeled nucleic acids is produced.

2. The method according to claim 1, wherein the set of primers comprises at least 50 primers selected from SEQ ID NOS:

01-1372.

3. The method according to claim 1, wherein the labeled nucleic acids are first strand cDNA.

4. The method according to claim 1, wherein the generating step comprises at least one amplification step.

5. A method of analyzing the differences in the RNA profiles between a plurality of different physiological sources, said method comprising:

obtaining a sample of ribonucleic acids from each of the distinct physiological sources;

generating a population of labeled nucleic acids for each of the different physiological sources by using a set of primers comprising at least 20 primers selected from SEO ID NOS;

01-1372; and

comparing the populations of labeled nucleic acids for each physiological source to identify differences in the populations.

6. The method according to claim 5, wherein the comparing step comprises

hybridizing the labeled nucleic acids for each of the distinct physiological sources to an array of probe nucleic acids stably associated with the surface of a substrate to produce a hybridization pattern for each of the different physiological sources;

and

comparing the hybridization patterns for each of the different physiological sources.

7. The method according to claim 5, wherein the labeled nucleic acids are first strand cDNA.

8. The method according to claim 5, wherein the generating step comprises an amplification step.

9. A method of analyzing the differential expression of genes in a plurality of distinct cell types, said method comprising:

obtaining a sample of distinct ribonucleic acids from each of the distinct cell types;

generating a population of labeled nucleic acids for each of the distinct cell types by using a set of primers comprising at least 20 primers selected from SEQ ID NOS;

01-1372;

hybridizing the labeled nucleic acids for each of the distinct cell types to an array of probe nucleic acids stably associated with the surface of a substrate to produce a hybridization pattern for each of the distinct cell types; and

comparing the hybridization patterns for each of the distinct cell types.

10. The method according to claim 9, wherein the labeled nucleic acids are first strand cDNA.

11. The method according to claim 9, wherein the generating step comprises an amplification step.

12. The method according to claim 9, wherein the number of primers in the set ranges from 20 to 1372.

13. The method according to claim 10, wherein the number of primers in the set does not vary by more than 50% from the number of probes bound to the substrate.

14. The method according to claim 9, wherein the distinct cell types share a common genome.

15. A method of analyzing the differential expression of genes in at least two different cell types sharing a common genome, said method comprising:

obtaining a population of distinct messenger ribonucleic acids from each of the different cell types;

generating a population of labeled nucleic acids for each of the different cell types by using a set of primers comprising at least 20 genes selected from SEQ ID NOS;

01-1372, wherein the number of gene specific primers in the set ranges from 20 to 1372,

hybridizing the labeled nucleic acids for each of the different cell types to an array of probe nucleic acids stably associated with the surface of a substrate to produce a hybridization pattern for each of the different cell types; and

comparing the hybridization patterns for each of the different cell types.

16. The method according to claim 15, wherein the labeled nucleic acids are first strand cDNA.

17. The method according to claim 15, wherein the generating step comprises an amplification step.