IGEM:VGEM/2007/Notebook/2007-8-1

Expect synthetic DNA from GENEART for melanin project

• Making Minimal Medium:
  • Add 495mL of distilled water to 3.0g Na2HPO4, 1.5g KH2PO4, 0.25g NaCl, 0.5g NH4Cl
  • Repeat above step in another flask with the addition of 7g of agar
  • Adjust pH to 7.4
  • Autoclave at 121oC

• • Add 3.02g MgSO4 powder to 25mL of water to make 1M solution
  • Add 8g glucose to 32mL water
  • Autoclave

• • Add 1.0mL 1M MgSO4 solution to broth and to agar solutions
  • Add 5.0mL of 20% glucose solution to broth and agar solutions

• • After cooling, pour agar solution into plates

Extracting and plating K12 Cells:

• • Extract K12 cells from glass vial by heating the tip over a bunson burner flame then cold shocking it
  • Incubate tube and plates at 37oC for 24hr