• 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

Protocols

Molecular Biology

Products and stock preparation

• Primer preparation: initial dilution of ordered primers.
• Competent Cells: prepare cells for plasmid uptake.
• Competent Cells. Protocol II (Inoue method).
• Glycerol stock: stock transformed cells at -80°C.
• Agar plate preparation: preparing agar plates for cell cultures.
• Autoclave: to sterilize solutions and glassware.

Cloning, assembly, and mutations

• Transformation: introduce foreign plasmid into competent cells.
• Plating: plate transformed cells
• Liquid cultures: when cells have grown on plates, put them in liquid cultures
• Gibson assembly.
• Linear template- TetR.
• tetR Overlap Extension PCR: induce specific mutations on tetR linear template.
• tetR Site-specific mutagenesis: induce site-specific mutations on a plasmid containing tetR.
• Agarose gel electrophoresis: DNA samples analysis (can be followed by band purification).
• Colony PCR: Analyze the plated transformed cells, to see if Gibson assembly worked

DNA recovery

• Plasmid preparation - Miniprep: recover plasmids from a bacterial culture.
• Gel purification: recover DNA separated by electrophoresis

Biochemistry

• IPTG test: test expression of a gene downstream from Plac

Microfluidics

• PDMS two layer device fabrication
• Cast microfabrication for PDMS chip
• MITOMI: Protein – DNA interactions
• Chemostat cell culture
• Klenow dsDNA synthesis