User:NYOUSHA Yousefi/Notebook/Protein Engineering/2013/01/29

Entry title

Preparing protein solutions and lyophilization

Objective 1

Eight different protein solutions were prepared for lyophilization

Procedure 1

1. 0.2g D-manitol was added to four set of tubes and 0.2g P-sorbitol was added to another set of four tubes, each tube recieved 0.2g of sugar
2. 0.1g of Polyethylene Glycol (PEG), M.W. 20,000 was added to all tubes
3. 10ml of four set of buffers including phosphate, tris, citrate, and acetate were added to tubes in an order that each four set of tubes received only one of these buffers
4. The tubes were vortexed to dissolve PEG into the buffer and sugar solution
5. Finally, the tubes were placed in -80 degree C nitrogen to freeze and then they were placed in the vacuum for lyophilization

Objective 2

Culturing PQE-80 antibiotic resistant bacteria

Procedure 2

1. 5ml of LB medium was added to two set of sterile tubes by 10 ml pipets
2. 5ul of Ampicillin was added to both test tubes
3. The frozen cells of PQE-80 bacteria in glycerol were taken out of the -80 degree C fridge
4. Some PQE-80 bacteria cells were added to these LB mediums under sterile conditions
5. PQE-80 bacteria were also added to four erlyn meyer flasks containing LB media and 35ul Ampicillin
6. The tubes and flasks were placed in the incubator/shaker at 37 degree C for 24h