Continue preparing for the MBP-intein expression. Continue the concentration of the purified GFP. Reproduce results from 12/07/11 with gold nanoparticles and refolding buffer.
Prep for MBP-intein expression
- Dry cycle in autoclave of various sizes of centrifuge tubes for expression next week.
- Combine 25g LB with 2g of D-glucose and 1L of distilled H2O in a 2800mL flask. (prepare 2)
- Combine 0.875g LB with 0.07g D-glucose and 35mL of distilled H2O in a 250mL flask. (prepare 2)
- Autoclave all of this media in a liquid cycle.
Studying the Effect of Refolding Buffer (repeat procedure from 12/07/11)
- Mix 100μL of 2.76mM HAuCl4 and 100μL 15.06μM BSA with 800μL distilled water in a microcentrifuge tube.
- Place the tube on a heat block at 80°C for three hours.
- Add 500μL 50mM Tris, pH 7.5.
- Remove from heat after about 5 min. Observe.
Continuation of Concentration
- Add more binding buffer (20mM sodium phosphate, 0.5M NaCl, 20mM imidazole) to concentrator, and continue concentrating the purified GFP to about 10mL.
- Sterile filter the concentrated GFP and store at 4°C.
- Purple protein fibers formed before the solution was even removed from the heat block.