13 August 2007

Preparation of Stock Solutions

• 1x 1 litre of 2xYT medium in Duran bottle
• 3x 1 litre of 2xYT medium in conical flask
  • Sent to autoclave
• Solution A for preparation of vesicles

Transformation of Biobricks

1. 7 biobrick were transformed into XL1-Blue or BL21(DE3)
2. Cells were grown for 1 hour and plated
3. Plates were incubated at 37°C overnight

• Transformed in XL1-Blue
  • BBa_J5528 [pBad-GFP] (re-clone)
  • BBa_E0240 [GFP]
  • BBa_I13507 [RFP]
  • BBa_T9002 [ptet-luxR-pLux-GFP]
  • BBa_B0034 [RBS]
• Transformed in BL21(DE3)
  • BBa_I13522 [ptet-GFP]
  • BBa_E7104 [T7-GFP]

Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Electrophoresis Electroporation] in the general protocols page

Pilot Preparation of Vesicles

1. Followed the desiccation and suspension stages of the vesicle preparation protocol
2. One 100ml beaker was prepared with 125μl of DOPC in 50ml of mineral oil
3. The suspension was sonicated for 30mins and left overnight in a 27°C incubator