• Introduction
  • Home
  • The Team
  • Photo Gallery
• Our Project
  • Results Summary
  • Selection System
  • In-Vitro Characterization
  • In-Vivo Characterization
  • Microfluidics
  • Data
  • Attributions
• Tools
  • Gibson Assembly
  • MITOMI
• Notebook
  • Protocols
  • May
  • June
  • July
  • August
  • September
  • October
• Considerations
  • Human Practices
  • Safety
• Acknowledgements
  • Sponsors
  • Partners

Todo

Microfluidic Chips

• Continue alignment training
• Run experiments on "chemostat" chip to check design
• Adapt design of "chemostat" chip for e-coli

Preparing the parts

• Sequence the lysis cassette
• Double-check lysis cassette sequence

All the parts are verified, we can now assemble them!

Assembly

• Research plasmid backbones. Is there one that already contains Tet-repressed LacI or GFP?
• Design Gibson primers to assemble the three different plasmids.
• Receive said primers
• Determine which sequence on LacI-plasmid and Lysis-plasmid should be used to create the "mega-plasmid".
• Think of new assemblies we want to make (pTet with RFP, for example)

Specifically:

• Run PCRs and gels on existing sequences (plasmid backbone, TetR gene, RFP)

TetR mutants

• determine required sequences
• order primers
• ext. PCR for MITOMI

MITOMI

• repeat MITOMI experiments for practice

• order one-off sequences for tetR MITOMI
• de Brujin experiment on TetR (will also yield PWM)
• determine position weight matrix for TetR, compare with de Brujin results
• determine position weight matrix for TetR mutants

Microfluidics and chemostat chip

• Continue alignment training

• Repeat experiments to check design
• Grow E. Coli from spotted arrays
• [No microfluidics] Setup a plate and test tween concentrations
• Determine growth rate as function of tween concentration (say 0.075% +- 0.7, as many increments as will fit on the chip)

Wiki

Protocols

• Describe cell cultures in miniprep protocol
• Write a new protocol!
• Upload "chemostat" protocols

General

• Write-up team presentation
• Upload our initial research about transcription factors

Clean room

• Order lab notebook
• Order storage box