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 a plastic floating clamp.
|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

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SpectraPor Membrane Dialysis
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.
Pierce Slide-A-Lyzer Dialysis
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.