Yu:Dialysis: Difference between revisions
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|4. Rinse thoroughly in a large volume of distilled H<sub>2</sub>O. | |4. Rinse thoroughly in a large volume of distilled H<sub>2</sub>O. | ||
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|5. Carefully tie off one end of the tubing with a double-knot. | |5. Carefully tie off one end of the tubing with a double-knot. Clamp directly above the knot. | ||
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|6. Add solution to be dialyzed by pipetting. | |6. Add solution to be dialyzed by pipetting. | ||
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|7. Clamp off the open end with | |7. Clamp off the open end with two plastic floating clamps. | ||
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|8. Suspend in the dialysis solution in a large beaker or graduated cylinder, depending on the length of the dialysis tubing. | |8. Suspend in the dialysis solution in a large beaker or graduated cylinder, depending on the length of the dialysis tubing. |
Latest revision as of 10:32, 22 May 2010
1. Cut an appropriate length of dialysis tubing. You will need 5cm for tying, plus 5cm tubing per 1mL. |
2. Soak in a large volume of distilled H2O for 30 minutes |
3. Soak in a large volume of 2% Sodium Bicarbonate (w/v), 1mM EDTA at 60°C for 30 minutes |
4. Rinse thoroughly in a large volume of distilled H2O. |
5. Carefully tie off one end of the tubing with a double-knot. Clamp directly above the knot. |
6. Add solution to be dialyzed by pipetting. |
7. Clamp off the open end with two plastic floating clamps. |
8. Suspend in the dialysis solution in a large beaker or graduated cylinder, depending on the length of the dialysis tubing. |
9. Change dialysis solution as needed. |
10. When dialysis is complete, carefully pinch off the tubing under the clamp. Remove the clamp carefully - note that the tubing will break where it was clamped if you are not supporting the tubing. |
11. Carefully decant the dialyzed solution by pipetting or by pouring. |
1. Soak cassette in dialysis buffer for 30 seconds. |
2. Carefully draw sample into a syringe with an 18-21 gauge needle (1-2"). |
3. Insert needle into corner gasket and inject sample without contacting the dialysis membrane. Withdraw any air in the membrane. |
4. Mark the corner gasket used with a permanent marker. Do not reuse entry points. |
5. Suspend in the dialysis solution in a large beaker. Change solution as needed. |
6. When dialysis is complete, draw back a new syringe to obtain a volume of air equivalent to sample volume. Insert needle into an unused corner gasket and separate dialysis membranes by injecting air. Draw off sample. |