IGEM:Harvard/2006/ProteinCoupling

provided by Dr. Shih

Generate a fresh aliquot of 0.2M EDC (dissolve 10 mg at a time).

Positive control: 10 uL unwashed optilink (non-magnetic) beads (10% suspension) + 10 uL 1 M MES pH 6.0 + 100 uL 100 uM aminomethylfluorescein + 20 uL 0.2M EDC (freshly dissolved)

Negative control: 10 uL unwashed optilink (non-magnetic) beads (10% suspension) + 10 uL 1 M MES pH 6.0 + 100 uL 100 uM aminomethylfluorescein + 20 uL water

Incubate 15 min at room temp. Spin in microfuge 15000 rcf 1 min. Remove the rest of the supernatant carefully, discard. Resuspend pellet in 100 uL 0.1M HEPES pH 7.5. Record fluorescence of the pellets on UV box using digital camera (same setup used to take pictures of ethidium bromide stained gels).