Methods for repairing degraded DNA

Patent metadata

Field	Value
Publication number	US-9873906-B2
Application number	US-90010410-A
Country	US
Kind code	B2
Filing date	Oct 7, 2010
Priority date	Jul 14, 2004
Publication date	Jan 23, 2018
Grant date	Jan 23, 2018

Abstract

The present invention provides methods and kits for repair of degraded DNA which may then be used as a template for efficient amplification by a number of different amplification reactions. The method relies upon a series of enzymatic activities provided by DNA repair enzymes.

First claim

What is claimed is:

1. A kit comprising a purified endonuclease, a purified DNA N-glycosylase, a purified AP lyase, a purified DNA polymerase, a purified 3′-diesterase, a purified polynucleotide kinase, a purified large Klenow fragment, a purified DNA ligase, a mixture of random primers comprising an equal part mix of hexamers, heptamers, and octamers with thiophosphate linkages for the last two 3′ nucleotides and a mixture of deoxynucleotide triphosphates, wherein the kit optionally further includes a highly processive strand displacing polymerase.

2. The kit of claim 1 further comprising irradiated and non-irradiated DNA controls.

3. The kit of claim 1 further comprising pyrophosphatase.

4. The kit of claim 1 further comprising φ29 polymerase.

5. The kit of claim 1 further comprising dithiothreitol (DTT).

6. The kit of claim 1 further comprising bovine serum albumin (BSA)-acetylated.

7. The kit of claim 1 further comprising glycerol.

Assignees

• Eshoo Mark W
• Ibis Biosciences Inc

Inventors

• Eshoo Mark W

Classifications

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q2521/531

  Glycosylase · CPC title

• C12Q2521/501

  Ligase · CPC title

• C12Q2521/514

  Mismatch repair protein · CPC title

• C12Q2521/301

  Endonuclease · CPC title