Hybridization device, methods, and system using mixing beads
View Patent ↗A method, device and system for hybridizing a target oligonucleotide to at least one array comprising a plurality of mixing beads are provided. A target solution is mixed by agitating the mixing beads while the target oligonucleotides are hybridizing to the complementary probes on the array. In another embodiment, a permeable barrier contains the mixing beads, thereby preventing them from contacting the array surface.
1. A method for hybridizing a target nucleic acid to a probe on an array, the method comprising:
providing a first substrate, wherein the first substrate comprises a support member, and wherein the array is attached to the support member;
providing a second substrate, wherein the second substrate comprises a container having a sub-chamber comprising a permeable barrier;
providing mixing beads, wherein the mixing beads are contained within the permeable barrier in the sub-chamber of the container such that a direct contact between the mixing beads and the surface of the array is prevented when the array is in the container;
adding a solution to the container, wherein the solution includes the target nucleic acid;
dipping the array into the solution; and hybridizing the target nucleic acid to the probe, wherein the hybridization includes agitation of the mixing beads, and wherein the agitation of the mixing beads additionally mixes the solution.
2. The method according to claim 1 , wherein the mixing beads are about 20 to about 400 microns in diameter.
3. The method according to claim 2 , wherein the permeable barrier is a mesh.
4. The method according to claim 1 , wherein the mixing beads are magnetic.
5. The method according to claim 4 , wherein the agitation of the mixing beads comprises applying a magnetic field to the mixing beads, and wherein said applying of the magnetic field to the mixing beads effectuates mixing of the solution.
6. The method according to claim 5 , wherein said applying of the magnetic field further prevents the contact between the mixing beads and the array.
7. The method according to claim 4 , wherein the mixing beads are made from stainless steel and are spherical in shape.
8. The method according to claim 1 , wherein the support member is a peg.
9. The method according to claim 8 , wherein the second substrate is a hybridization plate.
10. The method according to claim 1 , wherein the first substrate further comprises a plurality of support members, wherein a plurality of arrays is attached to the plurality of support members, and wherein the second substrate further comprises a plurality of containers.
11. The method according to claim 1 , wherein the mixing beads are about 50 to about 150 microns in diameter.
12. The method according to claim 1 , wherein the mixing beads are about 90 microns in diameter.
13. The method according to claim 1 , wherein the permeable barrier is a disposable mesh insert comprising spacers.
14. The method according to claim 1 , wherein the permeable barrier is a mesh insert that encloses the mixing beads.
15. The method according to claim 1 , wherein the container includes a surface comprising features that cause the mixing beads to move along a Z-axis of the container during the agitation step.
16. The method according to claim 15 , wherein the surface comprising features is a curved surface or includes channels, ridges, pockets, or bumps.
17. The method according to claim 15 , wherein the container is a well and a surface comprising features is located on the bottom of the well.
18. The method according to claim 1 , wherein the volume of the solution added to the container is about 20 microliters to about 100 microliters.
19. The method according to claim 1 , wherein the volume of the solution added to the container is about 40 microliters to about 80 microliters.
20. The method according to claim 1 , wherein the volume of the solution added to the container is about 60 microliters.