Methods and materials for optimization of electronic transportation and hybridization reactions
View Patent ↗Methods for the transport and hybridization of a nucleic acid on an electrode device by providing a low conductivity buffer with a reducing agent to the device. The low conductivity buffer may also contain a zwitterion. A current and voltage is applied to a location of the device to effect electrophoretic transportation of the nucleic acid towards the location. The nucleic acid is then hybridized to a nucleic probed located at the location. The reducing agent in the low conductivity buffer may also be acting as a chaotropic agent.
1. A method for the transport of a nucleic acid to an electrode site comprising the steps of:
providing a nucleic acid and a low conductivity, zwitterionic buffer with a reducing agent to an active electronic system comprising an electrode;
applying current and voltage to the electrode to effect electrophoretic transportation of the nucleic acid towards the electrode; and
hybridizing the nucleic acid with a probe located at the electrode, whereby the local pH above the electrode is below the pH of the buffer at its isoelectric point.
2. The method of claim 1 , wherein the reducing agent is α-thioglycerol.
3. The method of claim 1 , wherein the reducing agent is dithiotreitol.
4. The method of claim 1 , wherein the reducing agent is β-mercaptoethanol.
5. The method of claim 1 , wherein a concentration of the reducing agent is about 25-500 mM.
6. The method of claim 1 , wherein a concentration of the reducing agent is about 25 mM.
7. The method of claim 1 , wherein a concentration of the reducing agent is about 125 mM.
8. The method of claim 1 , wherein a concentration of the reducing agent is about 150 mM.
9. The method of claim 1 , wherein a concentration of the reducing agent is about 250 mM.
10. The method of claim 1 , wherein a concentration of the reducing agent is about 500 mM.
11. The method of claim 1 , wherein the low conductivity, zwitterionic buffer is histidine.
12. The method of claim 11 , wherein the low conductivity, zwitterionic buffer further comprises an additional zwitterion.
13. The method of claim 1 , wherein the low conductivity, zwitterionic buffer contains L-histidine.
14. The method of claim 1 , wherein the low conductivity, zwitterionic buffer contains D-histidine.
15. The method of claim 1 , wherein the low conductivity, zwitterionic buffer contains cysteine.
16. The method of claim 1 , wherein the low conductivity, zwitterionic buffer contains alanine.
17. The method of claim 1 , wherein the low conductivity, zwitterionic buffer contains lysine.
18. The method of claim 1 , wherein the low conductivity, zwitterionic buffer contains glutamic acid.
19. The method of claim 1 , wherein the low conductivity, zwitterionic buffer contains γ-aminobutyric acid.