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 == | == plasmid & GFP-double terminator's Ligation & Transformation == | ||
- | # | + | #added 2 uL TE into plasmid solvant and GFP-double terminator solvant |
- | # | + | #mixed the samples |
- | #added 5 uL Mighty mix | + | #added 5 uL Mighty mix |
- | #incubated at 16C for 30min | + | #incubated at 16C for 30min |
- | #added the sample to 100 uL competent cell | + | #added the sample to 100 uL competent cell |
- | #incubated at 0C for 30min | + | #incubated at 0C for 30min |
- | #heatshocked at 42C for 60sec | + | #heatshocked at 42C for 60sec |
- | #incubated at 0C for 5min. | + | #incubated at 0C for 5min |
+ | #added sample to 400 uL LB | ||
+ | #incubated at 37C for 2 hours | ||
+ | #plated the sample on LBA medium | ||
+ | #incubated at 37C | ||
+ | |||
+ | == 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
- added 2 uL TE into plasmid solvant and GFP-double terminator solvant
- mixed the samples
- added 5 uL Mighty mix
- incubated at 16C for 30min
- added the sample to 100 uL competent cell
- incubated at 0C for 30min
- heatshocked at 42C for 60sec
- incubated at 0C for 5min
- added sample to 400 uL LB
- incubated at 37C for 2 hours
- plated the sample on LBA medium
- incubated at 37C