User:NYOUSHA Yousefi/Notebook/Protein Engineering/2013/01/22

Entry title

Culturing antibiotic resistant bacteria producing myoglobin and hemoglobin

Objective 1

Ampicillin and Kenamycin were the antibiotics used for culturing

Procedure 1

1. 5ml of LB medium was added to two set of sterile tubes by 10 ml pipets
2. 5ul of Ampicillin was added to one tube and Kenamycin was added to the other tube
3. The frozen cells in glycerol were taken out of the -80 degree C fridge
4. Bacteria expressing Myoglobin were added to the media tube containing Ampicillin because it has Ampicillin resistance
5. PQE-80 bacteria were added to media tube containing Kenamycin becasue it has Kenamycin resistance
6. The tubes were placed in the incubator/shaker at 37 degree C for 24h

Objective 2

Lyophilization of 0.0058g Myoglobin, 20ml PH 7 phosphate, 0.4961 mg KCL sample

Procedure 2

1. The tube of the sample was placed in liquid nitrogen for 15 min to completely freeze it
2. When all the liquid of the sample was solidified, the sample were place under vacuum to make the ice to sublimate into vapor water directly and to leave the sample
3. The vacuum was allowed to run over night for 24h