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Label your tubes as instructed (1-8 or all 8 one number). Ask your teacher!
Prepare a lysozyme and salt solution for "Experiment Station."
You will dissolve the lysozyme in the acetate buffer.
1. Bring tour team's Erlenmeyer flask or medicine portion cup to the stock station.
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. While one teammate is getting the buffer another teammate can be setting up the array tubes numbering them 1 through 8 near the top of the tube.
. Measure out exactly 10 ml of the buffer to 15-ml conical tube. Use a dropper pipette to remove and excess volume. Put the excess volume in the waste beaker (Marked).
4. Pour the 10ml of buffer solution from the measuring tube into the Erlenmeyer flask and return with the flask to your experiment station.
5. Place a clean, dry powder funnel in the Erlenmeyer flask.
6. To add lysozyme (0.4), hold a gelatin capsule vertically and tap gently to get most of the powder to bottom carefully pull off the cap (it helps to turn or twist the cap first) and pour the lysozyme into the funnel. Tap the capsule and funnel then recap the capsule. Take care not to create or inhale and lysozyme dust.
7. To dissolve the lysozyme let it sit on the surface of the solution. (The less lysozyme on the glass, the better.) It may help to tap the flask on a padded surface to start the dissolving process. After the lysozyme has dissolved (5-10 min.) roll or swirl the flask to incorporate any clinging powder. Now that the lysozyme has been added it is important to avoid creating bubbles unnecessarily. Bubbles that form in protein solutions denature the protein (not desirable.). Now you have a buffer solution supersaturated with lysozyme at a level of 40mg/ml. It will take several minutes for proteins to dissolve.
Add the salt to the dissolved lysozyme in buffer
8. Tap down the salt crystals in the snap-cap tube.
. Carefully, open the snap-cap tube
10. Very slowly add the salt to the Erlenmeyer flask while swirling the flask but avoid splashing the liquid. Do this by tapping several salt crystals each time. Stop adding salt it begins to accumulate on the bottom of the flask but keep swirling.
11. Continue slowly adding salt after the accumulation has dissolved. This is to keep the concentration from becoming too strong in a local area of the solution. Too much salt could cause the liquid to turn cloudy; meaning protein is quickly coming out of solution.
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