Gel-Filteration Prelab

Gel-Filteration Prelab - column Continue to add until beads...

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Pre Lab Gel-Filtration Chromatography Purpose: To pack a gel filtration column and separate a mixture of three colored compounds on the basis of differences in molecular weight. Techniques: Do not punctuate the inner bottom surface of the column. Do not let the beads dry. Steps: Degas the hydrated Sephadex G-100 for 30 minutes. In the meantime twist off the bottom end if it is a new column. Put 5 mL of water in the column. Make 50mM TrisCl pH 8 buffer. Swirl and mix the bead and the buffer to form slurry. Open the lid and collect the liquid. Pour the slurry into the column before all the water is out. Use several mL of 50 mM TrisCl pH 8 to rinse the flask and pour it along the glass rod onto the
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Unformatted text preview: column. Continue to add until beads settle. Use to 5 times volume of 50 mM TrisCl pH8 to equilibrate the column. After volume passes through the resin close the small yellow lid. Level of buffer should be just above the resin bed. Read the absorbance value of blue dextran, using 50mM TrisCl buffer as a blank. Use 3 column volume of 20% entonlol to wash the column. Then leave 10 mL above the bed, cap the column, close the outlet, gently invert the column to generate slurry and pour into container. Rinse empty column with water the D water and return it....
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