• 9 am to 2:30pm:We take out 25 cryotubes incubated at 37 degrees C from Monday night:
  • 1 x Special one E7
  • 4 x GFP
  • 4 x RBS 0032
  • 4 x RBS 0034
  • 4 x Fe promoter
  • 4 x T7 promoter
  • 4x supD
• For all the 25 samples, we then centrifuge (for 70 mins in total), do the miniprep (for 120 mins in total) and digestion with EcoRI and PstI (120 mins incubation afterwards). The digestion is for gel electrophoresis, which is run after lunchtime. The detailed amount (in ul) for each digestion mixture is:

EcoRI: 0.5
PstI: 0.5
BSA: 0.2
EcoRI buffer: 2
DNA: 1
DI water: 5.8
Total: 10 ul

• 2:30 pm - 4:30pm: sequential digestion for pLacI insert and empty plasmid vector extracted from RBS 34.