D. Ficus Transformation Preperations.

 This week has spent planning and prepping for the transformation of D. Ficus. 

4.29

  • TGY media for plates was too concentrated agar wise, remade to 500ml from 250ml and re-autoclaved.
4.30

  • Poured 14 TGY plates for future inoculation. 
  • 2 TGY plates were inoculated w. D. Ficus
    • cells were from February, may not grow. Need to observe. 
  • Made 5x flask of 50ml of LB broth for future E. Coli inoculations. 
    • .1ml/flask of amp(25mg/ml)
5.1
  • Inoculated 2x 20ml TGY flask with D. Ficus after positive gram stain results
  • Inoculated 2x 50ml LB flask with E.Coli, added 100ul of 25mg/ml amp.
  • Discussed further plans of plasmid extraction and component cell protocols and material request.
  • Planned for transformation next week.
5.2
  •     Gram stained all cultures, both E. coli and D. Ficus
    • All good results
  • Took OD values of chosen cultures. 
    • E. Coli 2 OD 600 = 2.10
    • D. Ficus 2 OD 600 = 2.93
  • Held Biotech Club Meeting
  • Prepped for Plasmid Extractions and Component Cell Protocol for 5.3.24 had to be moved due to meeting. 
5.3
  •    Performed 2 plasmid extraction on E. Coli, 
    • Sample 1 - 179ng/uL
    • Sample 2 - 146ng/uL
  • Performed 2 component cell protocols on D. Ficus, set in -80 to store.
  • Planned for gel and possible transformation on Monday. 

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