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Team:Tokyo Metropolitan/Notebook/S29
From 2011.igem.org
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oriT - GFP for RP4 | oriT - GFP for RP4 | ||
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| + | ----- | ||
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| + | @colony PCR oriTr-GFP-oriTr | ||
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| + | @preculture oriTr-GFP-oriTr | ||
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| + | = -Targeting- = | ||
| + | Colony PCR | ||
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| + | Colony PCR J23100-P0140(J61002)-pSB1C3,I13522-pSB1C3 and AHL inducible Device①②(pSB1C3)-E0240.But failure. | ||
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| + | electrophoresis | ||
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| + | Anti-killer Device⑥(pSB1C3),Anti-killer Device⑦(pSB1C3) yesterday emzimed and find binds. | ||
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| + | preculture | ||
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| + | J23100-P0140(J61002) Master Plate No.9,I13522(pSB1C3) Master Plate No.2,AHL inducible Device①-E0240 Master Plate No.1,AHL | ||
| + | inducible Device②-E0240 Master Plate No.2 | ||
Latest revision as of 14:10, 5 October 2011
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-Visualization-
-Plasmid extraction
Promoter - suppressed expression system(Am)
-Reining
Above samples
-Make competent cells
F+ and RP4
Cultured to OD600:0.6
Harvested
Suspended with 0.1 M CaCl2
Incubated on ice for 10 min
Harvested and abandoned supernatant
Suspended 0.1 M CaCl2
-Transformation
Promoter - suppressed expression system for F+
oriT - GFP for RP4
@colony PCR oriTr-GFP-oriTr
@preculture oriTr-GFP-oriTr
-Targeting-
Colony PCR
Colony PCR J23100-P0140(J61002)-pSB1C3,I13522-pSB1C3 and AHL inducible Device①②(pSB1C3)-E0240.But failure.
electrophoresis
Anti-killer Device⑥(pSB1C3),Anti-killer Device⑦(pSB1C3) yesterday emzimed and find binds.
preculture
J23100-P0140(J61002) Master Plate No.9,I13522(pSB1C3) Master Plate No.2,AHL inducible Device①-E0240 Master Plate No.1,AHL inducible Device②-E0240 Master Plate No.2
