ABSTRACT
We will be using crystallization wells designed for the sitting
drop method.
1. Crystallization wells and sealing tape
2. Sodium acetate (1 M solution)
3. Sodium chloride (powder)
4. Lysozyme
5. Polyethylene glycol 300 (PEG 300), citrate, isopropanol,
bromophenol blue
6. Purified fluorescent protein from Experiment 9 (optional)
7. Reservoir solutions or ingredients for GFP crystalliza-
tion (optional)
Control solution 8% w/ v NaCl (0.8 g to make 10 mL) 1 mL of 1 M NaOAc H2O to 10 mL
Test solution 1 40% w/ v PEG 300 100 mM acetate 200 mM NaCl
Test solution 2 35% v/ v isopropanol 100 mM acetate
Each team will receive a vial of lysozyme and some solubilization buffer (the buffer is 0.02 M sodium acetate, pH 4.6). Dissolve your lysozyme in buffer as follows:
Team 1: Add 1 mL of solubilization buffer (lysozyme concentration: 20 mg/ mL)
Team 2: Add 0.8 mL of solubilization buffer (lysozyme concentration: 25 mg/ mL)
Team 3: Add 0.5 mL of solubilization buffer (lysozyme concentration: 40 mg/ mL)
Team 4: Add 0.4 mL of solubilization buffer (lysozyme
concentration: 50 mg/ mL)
Team 5: Add 0.2 mL of solubilization buffer (lysozyme
concentration: 100 mg/ mL)
More than 5 teams: Repeat from Team 1
e following is the procedure for each condition:
1. Select crystallization well.
