Team:HokkaidoU Japan/Notebook/September28

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(plasmid & GFP-double terminator's Ligation & Transformation)
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*glycerol-stock of E.coli with salmonella's BAC library vector
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*making competent cell of E.coli with SPI2
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*plasmid & GFP-double terminator's Ligation & Transformation
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*PCR of E.coli with T3SSsignal and of GFP-double terminator
== plasmid & GFP-double terminator's Ligation & Transformation ==
== plasmid & GFP-double terminator's Ligation & Transformation ==
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#先日凍結した2つのサンプルが乾燥して少なくなっていたので、2 uLのTEを加えて懸濁した。
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#added 2 uL TE into plasmid solvant and GFP-double terminator solvant
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#2つのサンプルを混合した。
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#mixed the samples
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#added 5 uL Mighty mix.
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#added 5 uL Mighty mix
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#incubated at 16C for 30min.
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#incubated at 16C for 30min
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#added the sample to 100 uL competent cell.
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#added the sample to 100 uL competent cell
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#incubated at 0C for 30min.
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#incubated at 0C for 30min
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#heatshocked at 42C for 60sec.
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#heatshocked at 42C for 60sec
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#incubated at 0C for 5min.
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#incubated at 0C for 5min
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#added sample to 400 uL LB
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#incubated at 37C for 2 hours
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#plated the sample on LBA medium
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#incubated at 37C
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== PCR of E.coli with T3SSsignal and of GFP-double terminator ==

Revision as of 14:34, 28 September 2010

  • 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
== PCR of E.coli with T3SSsignal and of GFP-double terminator ==