Buffers for electrophoresis and use thereof
View Patent ↗Various embodiments provide, for example, buffer compositions and/or sieving formulations useful in connection with electrophoresis instruments, such as capillary electrophoresis (CE) devices. In various embodiments, a buffer composition can include Bis-Tris, TAPS and/or TAPSO, and, optionally, a chelating agent, such as EDTA. Methods of separating samples containing bio-molecules, such as DNA or RNA, are also described.
1. A buffer composition for electrophoresis of nucleic acids, the buffer composition comprising:
(i) Bis-Tris
(ii) TAPS; and
(iii) a metal-chelating agent, wherein the pH of the buffer is no less than 7.5.
2. The composition of claim 1 , wherein said metal-chelating agent includes EDTA.
3. The composition of claim 2 , wherein the composition is substantially detergent-free.
4. The composition of claim 1 , wherein the composition is substantially detergent-free.
5. A buffer composition for electrophoresis of nucleic acids, the buffer composition comprising:
(i) at least one of TAPS, TAPSO, and Asparagine; and
(ii) a compound of the formula: [HO(CH 2 ) m ] 3 C-N[(CH 2 ) n-OH] 2,
wherein: m is an integer of from 1 to 3, and n is an integer of from 1 to 4; and further comprising a metal-chelating agent, and wherein the buffer composition has a pH of no less than 7.5.
6. The composition of claim 5 , wherein said metal-chelating agent is EDTA.
7. The composition of claim 5 , wherein the composition is substantially detergent-free.
8. The buffer composition of claim 5 comprising TAPS.
9. The buffer composition of claim 5 comprising TAPSO.
10. The buffer composition of claim 5 comprising Asparagine.
11. A resolving-gel composition for electrophoresis of nucleic acids, the composition comprising:
(a) an organic polymer,
(b) Bis-Tris,
(c) one or more members of the group TAPS, TAPSO, and Asparagine; and
(d) a metal-chelating agent,
wherein the organic polymer comprises a sieving component comprising a non-crosslinked acrylamide polymer, and a surface interaction component comprising one or more non-crosslinked polymers selected from the group consisting of N,N-disubstituted polyacrylamide and N-substituted polyacrylamide, and wherein the resolving-gel composition has a pH of no less than 7.5.
12. The composition of claim 11 , having a pH greater than 7.
13. The composition of claim 11 , comprising TAPSO.
14. The composition of claim 11 comprising Asparagine.
15. An apparatus for resolving samples, comprising:
an anodic buffer chamber;
a cathodic buffer chamber;
an electrophoretic channel extending between and communicating said anodic and cathodic buffer chambers; and
a continuous buffer system comprising Bis-Tris held in said anodic and cathodic buffer chambers and in said channel;
wherein said continuous buffer system further comprises one or both of TAPS and TAPSO, and wherein said continuous buffer system has a pH of no less than 7.5.
16. The apparatus of claim 15 , further comprising a sieving medium held in said channel.
17. The apparatus of claim 15 , further comprising an organic polymer, wherein the organic polymer comprises a sieving component comprising a non-crosslinked acrylamide polymer, and a surface interaction component comprising one or more non- crosslinked polymers selected from the group consisting of N,N-disubstituted polyacrylamide and N-substituted polyacrylamide.
18. A method of electrophoresis of nucleic acids, comprising:
adding a buffer into an electrophoretic channel, wherein said buffer comprises Bis-Tris, TAPS and a metal-chelating agent, and wherein said buffer has a pH of no less than 7.5;
adding a sample including nucleic acids to be analyzed into said channel; and
resolving said sample by electrophoresis, wherein said channel is defined by a capillary tube.
19. A method of electrophoresis of nucleic acids, comprising:
(a) adding a buffer into an electrophoretic channel, wherein said buffer comprises a compound of the formula: [HO(CH 2 )m] 3 C-N[(CH 2 ) n-OH] 2,
wherein m is an integer of from 1 to 3, and n is an integer of from 1 to 4, and
wherein the buffer further comprises TAPS or TAPSO and a metal-chelating agent;
(b) adding a sample including nucleic acids to be analyzed into said channel; and
(c) resolving said sample by electrophoresis, at a pH of no less than 7.5,
wherein said channel is defined by a capillary tube, and wherein at least a portion of the gel is at an ambient temperature during the application of the electromotive potential difference across the gel.
20. The method of claim 19 , wherein the metal-chelating agent is EDTA.
21. The method of claim 19 , wherein the buffer includes TAPS.