Team:HokkaidoU Japan/Notebook/September28

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  • glycerol-stock of E.coli with salmonella's BAC library vector
  • making competent cell of E.coli with SPI2
  • plasmid & GFP-double terminator's Ligation & Transformation
  • PCR of E.coli with T3SSsignal and of GFP-double terminator

plasmid & GFP-double terminator's Ligation & Transformation

  1. added 2 uL TE into plasmid solvant and GFP-double terminator solvant
  2. mixed the samples
  3. added 5 uL Mighty mix
  4. incubated at 16C for 30min
  5. added the sample to 100 uL competent cell
  6. incubated at 0C for 30min
  7. heatshocked at 42C for 60sec
  8. incubated at 0C for 5min
  9. added sample to 400 uL LB
  10. incubated at 37C for 2 hours
  11. plated the sample on LBA medium
  12. incubated at 37C