Production and use of normalized DNA libraries

• US 5,763,239 A
• Filed: 06/18/1996
• Issued: 06/09/1998
• Est. Priority Date: 06/18/1996
• Status: Expired due to Fees

• Current Assignee: Biotex Pharma Investments, LLC
• Original Assignee: Diversa Corporation (BASF SE)
• Inventors: Short, Jay M., Mathur, Eric J.
• Primary Examiner(s): Elliott, George C.
• Assistant Examiner(s): MCGARRY, SEAN
• Application Number:
  US08/665,565
• Time in Patent Office: 721 Days
• Field of Search: 435/91.2, 435/6, 435/172.1, 536/25.4, 536/25.42
• US Class Current: 435/6.18
• CPC Class Codes: C12N 15/1093 General methods of preparing gene libraries, not provided for in other subgroups
• CPC Class Codes: C12N 15/1096 cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
• CPC Class Codes: C12Q 1/6811 Selection methods for production or design of target specific oligonucleotides or binding molecules

Abstract

Disclosed is a process for forming a normalized genomic DNA library from an environmental sample by (a) isolating a genomic DNA population from the environmental sample; (b) analyzing the complexity of the genomic DNA population so isolated; (c) at least one of (i) amplifying the copy number of the DNA population so isolated and (ii) recovering a fraction of the isolated genomic DNA having a desired characteristic; and (d) normalizing the representation of various DNAs within the genomic DNA population so as to form a normalized library of genomic DNA from the environmental sample. Also disclosed is a normalized genomic DNA library formed from an environmental sample by the process.

Claims

1. A process for forming a normalized genomic DNA library from an environmental sample, which comprises the steps of:

(a) isolating a genomic DNA population from the environmental sample;

(b) analyzing the complexity of the genomic DNA population so isolated;

(c) at least one of the steps selected from the group consisting of (i) amplifying the copy number of the DNA population so isolated and (ii) recovering a fraction of the isolated genomic DNA having a desired characteristic; and

(d) normalizing the representation of various DNAs within the genomic DNA population so as to form a normalized library of genomic DNA from the environmental sample.

2. The process of claim 1 wherein step (c) consists of recovering a fraction of the isolated genomic DNA having a desired characteristic.

3. The process of claim 1 wherein step (c) consists of amplifying the copy number of the DNA population so isolated.

4. The process of claim 1 wherein the step of amplifying the genomic DNA precedes the normalizing step.

5. The process of claim 1 wherein the step of normalizing the genomic DNA precedes the amplifying step.

6. The process of claim 1 which comprises both the steps of (i) amplifying the copy number of the DNA population so isolated and (ii) recovering a fraction of the isolated genomic DNA having a desired characteristic.

7. A normalized genomic DNA library formed from from an environmental sample by a process comprising the steps of:

(a) isolating a genomic DNA population from the environmental sample;

(b) analyzing the complexity of the genomic DNA population so isolated;

(c) at least one of (i) amplifying the copy number of the DNA population so isolated and (ii) recovering a fraction of the isolated genomic DNA having a desired characteristic; and

(d) normalizing the representation of various DNAs within the genomic DNA population so as to form a normalized library of genomic DNA from the environmental sample.

8. The library of claim 7 wherein step (c) consists of the step of recovering a fraction of the isolated genomic DNA having a desired characteristic.

9. The library of claim 7 wherein step (c) consists of the step of amplifying the copy number of the DNA population so isolated.

10. The library of claim 7 wherein in the process of forming said library the step of amplifying the genomic DNA precedes the normalizing step.

11. The library of claim 7 wherein in the process of forming said library the step of normalizing the genomic DNA precedes the amplifying step.

12. The library of claim 7 wherein the process of forming said library comprises both the steps of (I) amplifying the copy number of the DNA population so isolated and (ii) recovering a fraction of the isolated genomic DNA having a desired characteristic.

13. A process for forming a normalized library of genomic gene clusters from an environmental sample which comprises

(a) isolating a genomic DNA population from the environmental sample;

(b) analyzing the complexity of the genomic DNA population so isolated;

(c) at least one of (i) amplifying the copy number of the DNA population so isolated and (ii) recovering a fraction of the isolated genomic DNA having a desired characteristic; and

(d) normalizing the representation of various DNAs within the genomic DNA population so as to form a normalized library of genomic DNA from the environmental sample.

14. A normalized library of genomic gene clusters formed from an environmental sample by a process comprising the steps of

(a) isolating a genomic DNA population from the environmental sample;

(b) analyzing the complexity of the genomic DNA population so isolated;

(c) at least one of (I) amplifying the copy number of the DNA population so isolated and (ii) recovering a fraction of the isolated genomic DNA having a desired characteristic; and

(d) normalizing the representation of various DNAs within the genomic DNA population so as to form a normalized library of genomic DNA from the environmental sample.