METHOD AND APPARATUS FOR ISOLATING AND PURIFYING BIO-MOLECULES

• US 20100326827A1
• Filed: 06/24/2009
• Published: 12/30/2010
• Est. Priority Date: 06/24/2009
• Status: Abandoned Application

• Current Assignee: Industrial Technology Research Institute
• Original Assignee: Industrial Technology Research Institute
• Inventors: LIN, Shang-Chi, LIN, Chia-Chen
• Application Number:
  US12/490,514
• Publication Number:
  US 20100326827A1
• Time in Patent Office: Days
• US Class Current: 204/461
• CPC Class Codes: G01N 27/44756 Apparatus specially adapted therefor

Abstract

Electrophoresis systems, apparatus, and methods for isolating or purifying a bio-molecule are provided. The method includes using an electrophoresis apparatus for isolating or purifying a bio-molecule, wherein the electrophoresis apparatus includes: a separation tube including a first section for containing buffer and a second section including an electrophoresis gel, and a collection tube including a solid phase, wherein the solid phase fully covers the cross-sectional area of the collection tube, and the collection tube is detachably connected to an end adjacent to the second section of the separation tube.

Claims

1. An electrophoresis apparatus for isolating or purifying bio-molecules comprising:

a separation tube comprising a first section for containing buffer and a second section including an electrophoresis gel, and

a collection tube comprising a solid phase, wherein the solid phase fully covers the cross-sectional area of the collection tube, and the collection tube is detachably connected to an end adjacent to the second section of the separation tube.

2. The electrophoresis apparatus in claim 1, wherein the bio-molecules comprise nucleic acids, DNA, RNA, protein, peptide, or a mixture thereof.

3. The electrophoresis apparatus in claim 1, wherein the bio-molecules are small nucleic acids, DNA, or RNA sample of 4-40 nucleotides.

4. The electrophoresis apparatus in claim 1, wherein the solid phase is a cationic solid phase.

5. The electrophoresis apparatus in claim 4, wherein the cationic solid phase comprises at least one bead, membrane, filter, or a mixture thereof.

6. The electrophoresis apparatus in claim 4, wherein the cationic solid phase comprises tertiary or quaternary amino groups.

7. The electrophoresis apparatus in claim 5, wherein the cationic solid phase comprises a filter having a pore size, wherein the water is extruded through the filter at a flow rate of between about 0.005 and about 1 mL/min/mm² under between 2000×

g and 10000×

g centrifugation.

8. The electrophoresis apparatus in claim 5, wherein the cationic solid phase comprises a filter having a pore size, wherein the water is extruded through the filter at a flow rate of between about 0.0177 and about 0.15 mL/min/mm² under between 2000×

g and 10000×

g centrifugation.

9. The electrophoresis apparatus in claim 1, wherein the solid phase comprises quaternary amine agarose beads, tertiary amine cellulose beads, tertiary amine filter, or quaternary amine filter.

10. The electrophoresis apparatus in claim 1, wherein the separation tube has a diameter of 0.1 cm to 2 cm.

11. The electrophoresis apparatus in claim 1, wherein the electrophoresis gel has a length of 0.5 cm to 20 cm.

12. A method for purifying or isolating bio-molecules comprising:

providing the electrophoresis apparatus as claimed in claim 1;

loading a sample comprising at least one bio-molecule to the first section of the separation tube;

applying an electric field by implementing at least one anode and at least one cathode to drive the at least one bio-molecule into the solid phase, wherein the electrophoresis apparatus is located between the at least one anode and at least one cathode;

detaching the collection tube from the separation tube, and

purifying the at least one bio-molecule isolated in the collection tube.

13. The method in claim 12, wherein the solid phase is soaked in water or a buffer.

14. The method in claim 12, further comprising monitoring the location of the at least one bio-molecule during application of the electric field.

15. The method in claim 14, wherein the monitoring comprises detecting UV light illumination, fluorescence, or monitoring the location of an electrophoresis indicator.

16. The method in claim 12, wherein the at least one bio-molecule comprises DNA, RNA, protein, peptide, or a mixture thereof.

17. The method in claim 12, wherein the at least one bio-molecule is a small DNA or RNA sample of 4-40 nucleotides.

18. The method in claim 12, wherein the solid phase is a cationic solid phase.

19. The method in claim 18, wherein the cationic solid phase comprises at least one bead, membrane, filter, or a mixture thereof.

20. The method in claim 19, wherein the cationic solid phase comprises a filter having a pore size, wherein the water is extruded through said filter at a flow rate of between about 0.005 and about 1 mL/min/mm² under between 2000×

g and 10000×

g centrifugation.

21. The method in claim 19, wherein the cationic solid phase comprises a filter having a pore size, wherein the water is extruded through said filter at a flow rate of between about 0.0177 and about 0.15 mL/min/mm² under between 2000×

g and 10000×

g centrifugation.

22. The method in claim 18, wherein the cationic solid phase comprises tertiary or quaternary amino groups.

23. The method in claim 12, wherein the solid phase comprises quaternary amine agarose beads, tertiary amine cellulose beads, tertiary amine filter, or quaternary amine filter.

24. The method in claim 12, further comprising replacing a new collection tube with the collection tube to separately collect different bio-molecules.

25. A kit for isolating or purifying bio-molecules, comprising:

the electrophoresis apparatus as claimed in claim 1;

a running buffer;

at least one elution or purification buffer, and

an user instruction.