Koeris/Notebook/2007-1-17

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Thoughts on biofilms

Linares et al.

Transformations

Standard heat shock protocol usable for any kind of origin. Also good for multi-plasmid transformations

Reagents necessary

  • Chemically competent cells (see here)
  • Plasmid prep >5ng/ul concentration as the lower limit is acceptable
  • SOC

Protocol

  1. Thaw competent cells on ice
  2. Add plasmid DNA to cell, stir with pipette tip, do NOT pipette up and down
  3. Incubate for 30min on ice
  4. Heat shock at 42 deg C for 30s
  5. Cool on ice for 5min
  6. Add 1ml SOC to 100ul competent cells
  7. Incubate at 37 deg C for 1h at 300rpm
  8. Spread plate 550ul on LB agar with the appropriate resistance markers