UNIVERSAL BIOLOGICAL SAMPLE PROCESSING

• US 20100137575A1
• Filed: 11/25/2009
• Published: 06/03/2010
• Est. Priority Date: 12/03/2008
• Status: Abandoned Application

• Current Assignee: Integrated Nano-Technologies, LLC
• Original Assignee: Integrated Nano-Technologies, LLC
• Inventors: Murante, Richard S., Connolly, D. Michael, DeBoer, Charles, Tannous, Vera
• Application Number:
  US12/626,294
• Publication Number:
  US 20100137575A1
• Time in Patent Office: Days
• US Class Current: 536/25.410
• CPC Class Codes: C12N 15/1003 Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2523/301 Sonication
• CPC Class Codes: C12Q 2523/303 Applying a physical force on a nucleic acid
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2563/143 Magnetism, e.g. magnetic label

Abstract

A method for preparing a sample by utilizing a mechanical force in the presence of a size stabilizer to break apart the sample to obtain nucleic acid molecules in a usable size range.

Claims

1. A method for extracting nucleic acid fragments comprising the steps of:

providing a biological sample;

suspending said biological sample in a suspension solution comprising a size stabilizer;

applying a mechanical force to said biological sample to extract nucleic acid molecules;

the application of said mechanical force in the presence of said size stabilizer results in said nucleic acid molecules being reduced to a median fragment size having a median number of base pairs, wherein at least 60% of said nucleic acid molecules have a number of base pairs that are within 50% of said median number of base pairs.

2. The method of claim 1 wherein said mechanical force is a shearing force.

3. The method of claim 1 wherein said mechanical force is sonic vibration.

4. The method of claim 3 wherein said sonic vibration has shearing frequencies between 10,000 Hz and 10 MHz.

5. The method of claim 1, wherein at least 75% of said nucleic acid molecules after the application of said mechanical force have between 200 and 500 base pairs.

6. The method of claim 1, wherein at least 75% of said nucleic acid molecules after the application of said mechanical force have more than 1000 base pairs.

7. The method of claim 1, wherein said suspension solution further comprises disrupting beads.

8. The method of claim 7, wherein said disrupting heads are glass beads.

9. The method of claim 1, wherein said size stabilizer comprises at least one compound selected from the group consisting of sodium phosphate, guanidinium hydrochloride, dextran sulfate sodium dodecyl sulfate, sodium dodceyl benzene sulfate, polyethyleneglycol, anionic surfactants, dipole aprotic solvents, dimethylsulfoxide, dimethyl formamide, dimethylacetamide, hexamethyl phosphoramide, tetramethylurea, kaotropic salt, poly vinyl alcohol, polyethylenimine, poly acrylic acid and other polymeric acids.

10. The method of claim 1, wherein said size stabilizer is selected from the group consisting of sodium dodecyl sulfate and sodium dodceyl benzene sulfate

11. The method of claim 1 further comprising the step of binding the nucleic acid molecules to magnetic nanoparticles.

12. The method of claim 11 further comprising the step of collected said magnetic nanoparticles and the bound nucleic acid molecules and washing magnetic nanoparticles and the bound nucleic acid molecules.

13. The method of claim 12 further comprising the step of eluting said nucleic acid molecule from said magnetic nanoparticles.

14. The method of claim 13 further comprising the step of providing an elution solution comprising alcohol to elute said nucleic acid molecule from said magnetic nanoparticles.

15. The method of claim 11 further comprising the step of applying a magnetic field to concentrate said magnetic nanoparticles and the bound nucleic acid molecules.

16. The method of claim 1, wherein said nucleic acid molecules is reduced to a median number of base pairs between 200 and 10,000 base pairs.

17. The method of claim 1, wherein said size stabilizer limits the minimum size of said nucleic acid molecules achievable by said shearing force to 150 base pairs.

18. The method of claim 1, wherein said biological sample is selected from the group consisting of bacterial cells, spores, viruses and biological tissue.

19. The method of claim 1, wherein said mechanical force is applied via a nebulizer or a homogenizer.

20. The method of claim 1, wherein at least 75% of said nucleic acid molecules have a number of base pairs that are within 50% of the number of base pairs as said median number of base pairs.