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Timeline



Week 26

- Preparing for take-off


  • Inventory of the lab supplies
  • Pouring LB Agar plates
  • Amplification of the pET-Duet-1 vector
  • Assessment of the safety requirements
  • Preparing stocks for antibiotics, glycerol, LB & MilliQ


Week 27

- The Clone Wars


Gibson Assembly:
  • Linearizing pET-Duet-1 for Gibson Assembly
  • Our first Gibson Assembly!

Traditional cloning:
  • Amplification of the pET-Duet-1 vector
  • Nde1 & Kpn1 digestion of the pET-Duet-1 vector (MCS-2) for traditional cloning


Week 28

- Et tu, pET-Duet?

Gibson Assembly:
  • Linearizing pET-Duet-1 for Gibson Assembly and debugging
  • Amplification of the inserts for traditional cloning
  • Xbal & Pstl digestion of the pET-Duet-1 vector (MCS-1) for traditional cloning

Traditional cloning:
  • Xba1 & Pst1 digestion of the inserts (MCS-1) for traditional cloning
  • Nde1 & Kpn1 digestion of the inserts (MCS-2) for traditional cloning
  • Ligation