Methods of preparing nucleic acid for detection

• US 20070042400A1
• Filed: 05/08/2006
• Published: 02/22/2007
• Est. Priority Date: 11/10/2003
• Status: Abandoned Application

• Current Assignee: Investigen, Inc.
• Original Assignee: Investigen, Inc.
• Inventors: Zwick, Michael, Choi, K., Mccue, Kent
• Application Number:
  US11/429,746
• Publication Number:
  US 20070042400A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12N 15/10 Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence

Abstract

Methods of preparing nucleic acid from polysaccharide-containing samples for detection by providing one or more glycosidases to the sample to degrade polysaccharides are provided. The nucleic acids can further be extracted from the sample. The method is particularly useful for detecting nucleic acid in samples with high starch content.

Claims

1. A method of preparing nucleic acid from a polysaccharide-containing sample, said method comprising:

providing one or more glycosidases to the polysaccharide-containing sample, thereby generating a glycosidase-treated sample.

2. The method of claim 1, further comprising extracting said nucleic acid from the glycosidase-treated sample.

3. A method of detecting nucleic acid in a polysaccharide-containing sample, comprising:

preparing the nucleic acid according to the method of claim 1;

and

detecting the nucleic acid.

4. The method of claim 1, wherein the one or more glycosidases includes a glycosidase selected from the group consisting of a glycoamylase, a debranching enzyme, a heterosaccharide degrading enzyme, and a non-glucose homosaccharide degrading enzyme.

6. The method of claim 4, wherein the glycoamylase is selected from the group consisting of an alpha-amylase, a beta-amylase, a glucan alpha 1,4-glucosidase, a glucan alpha 1,4-glucosidase, and a glucan alpha 1,6-glucosidase.

12. The method of claim 2, wherein said extracting includes providing an alcohol to the sample.

16. The method of claim 1, wherein the polysaccharide-containing sample comprises starch.

19. The method of claim 1, wherein the polysaccharide-containing sample includes one or more components selected from the group consisting of corn meal, soy flour, wheat flour, corn starch, corn chips, and maltodextrin.

20. The method of claim 1, further comprising removing the polysaccharides from the glycosidase-treated sample.

21. The method of claim 1, further comprising removing cellular components from the sample.

24. The method of claim 21, wherein the cellular components are removed by providing to the sample one or more salts selected from the group consisting of potassium acetate and sodium acetate, thereby precipitating said cellular components.

25. The method of claim 21, wherein said extracting includes applying the sample to a column.

40. The method of claim 3, wherein detecting the nucleic acid includes amplification thereof.

5. (canceled)

7-11. -11. (canceled)

13-15. -15. (canceled)

17-18. -18. (canceled)

22-23. -23. (canceled)

26. (canceled)

27. A kit for preparing nucleic acid from a sample comprising:

a) a glycosidase; and

b) instructions for using the kit.

28. The kit of claim 27, wherein the glycosidase is selected from the group consisting of a glycoamylase, an alpha-amylase, a beta-amylase, a glucan alpha 1,4-glucosidase, and a glucan alpha 1,6-glucosidase.

33. The kit of claim 27, further comprising one or more salts selected from the group consisting of potassium acetate and sodium acetate.

34. The kit of claim 27, further comprising sodium dodecyl sulfate (SDS).

35. The kit of claim 27, further comprising an alcohol.

37. The kit of claim 27, further comprising a column.

38. The kit of claim 37, wherein the column contains glass beads or glass wool.

29-32. -32. (canceled)

36. (canceled)

39. (canceled)