Multiple displacement amplification with blocker DNA

• US 20070092899A1
• Filed: 08/24/2006
• Published: 04/26/2007
• Est. Priority Date: 10/21/2005
• Status: Abandoned Application

• Current Assignee: Lawrence Livermore National Security LLC (Government of the United States of America)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Christian, Allen, Hara, Christine
• Application Number:
  US11/509,996
• Publication Number:
  US 20070092899A1
• Time in Patent Office: Days
• US Class Current: 435/6.18
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 2537/1373 Displacement by a nucleic acid
• CPC Class Codes: C12Q 2537/163 blocking probe

Abstract

A method of whole genome amplification of a nucleic acid sample comprising providing a starting template of the nucleic acid sample, bringing the template together with blocking DNA, bringing the template together with DNA polymerase, and multiple displacement amplification of the template. The method can be used with a starting template of less than 1 ng of the nucleic acid sample.

Claims

1. A method of amplifying a nucleic acid sample, comprising the steps of:

bringing the nucleic acid sample together with blocking DNA,

adding DNA polymerase, and

performing multiple displacement amplification.

2. The method of amplifying a nucleic acid sample of claim 1 wherein the method is adapted to amplify nucleic acid of a specific species of organism and said step of bringing the nucleic acid sample together with blocking DNA comprises bringing the nucleic acid sample together with blocking DNA from a species other than said specific species of organism.

3. The method of amplifying a nucleic acid sample of claim 1 wherein the method is adapted to amplify nucleic acid of a human species and said step of bringing the nucleic acid sample together with blocking DNA comprises bringing the nucleic acid sample together with blocking DNA from a species other than said human species.

4. The method of amplifying a nucleic acid sample of claim 1 wherein said step of bringing the nucleic acid sample together with blocking DNA comprises bringing the nucleic acid sample together with cross-linked salmon sperm DNA.

5. The method of amplifying a nucleic acid sample of claim 1 including performing polymerase chain reaction for further amplification.

6. A method of amplifying a nucleic acid sample, comprising the steps of:

providing a starting template of the nucleic acid sample,

bringing said template together with blocking DNA,

bringing said template together with DNA polymerase, and

multiple displacement amplification of said template.

7. The method of amplifying a nucleic acid sample of claim 6 wherein said step of providing a starting template comprises providing a starting template of less than 1 ng of the nucleic acid sample.

8. The method of amplifying a nucleic acid sample of claim 6 wherein the method is adapted to amplify nucleic acid of a specific species of organism and said step of bringing said template together with blocking DNA comprises bringing said template together with blocking DNA from a species other than said specific species of organism.

9. The method of amplifying a nucleic acid sample of claim 6 wherein the method is adapted to amplify nucleic acid of a human species and said step of bringing said template together with blocking DNA comprises bringing said template together with blocking DNA from a species other than said human specific species.

10. The method of amplifying a nucleic acid sample of claim 6 wherein said step of bringing said template together with blocking DNA, comprises bringing said template together with cross-linked salmon sperm DNA.

11. The method of amplifying a nucleic acid sample of claim 6 including performing polymerase chain reaction of said template for further amplification.

12. A method of whole genome amplification of a nucleic acid sample, comprising the steps of:

providing a starting template of the nucleic acid sample,

bringing said template together with blocking DNA,

bringing said template together with DNA polymerase, and

multiple displacement amplification of said template.

13. The method of whole genome amplification of a nucleic acid sample of claim 12 wherein said step of providing a starting template comprises providing a starting template of less than 1 ng of the nucleic acid sample.

14. The method of whole genome amplification of a nucleic acid sample of claim 12 wherein the method is adapted to amplify nucleic acid of a specific species of organism and said step of bringing said template together with blocking DNA comprises bringing said template together with blocking DNA from a species other than said specific species of organism.

15. The method of whole genome amplification of a nucleic acid sample of claim 12 wherein the method is adapted to amplify nucleic acid of a human species and said step of bringing said template together with blocking DNA comprises bringing said template together with blocking DNA from a species other than said human specific species.

16. The method of whole genome amplification of a nucleic acid sample of claim 12 wherein said step of bringing said template together with blocking DNA, comprises bringing said template together with cross-linked salmon sperm DNA.

17. The method of whole genome amplification of a nucleic acid sample of claim 12 including performing polymerase chain reaction of said template for further amplification.