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Team:HokkaidoU Japan/Notebook/Color R0040-S05192-B0015
From 2013.igem.org
Maestro E.coli
Notebook
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Start
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Get chromoprotein DNA
Great thanks to iGEM Uppsala! -
Transformation
S05192
1µL DNA to DH5α -
Colony PCR & Culture
S05192, B0015
We confirmed transformation was completely successful.
Cultured them for subsequent experiments. -
Mini-prep
S05192, B0015 -
PCR & PCR purification
S05192
100b up, 200b dn, KOD Plus NEO -
Digestion
Cut B0015 with EcoRI, XbaI (using 10xM buffer)
Cut S05192 with EcoRI, SpeI (using 10xM buffer) -
Antarctic phosphatase
B0015 -
Ligation
Ligate B0015 with S05192 -
Transformation
S05192-B0015
5µL DNA to DH5α -
ColonyPCR & Culture
S05192-B0015
We confirmed transformation was completely successful.
Cultured it for subsequent experiments. -
PCR
R0040
100b up, 200b dn, KOD Plus NEO -
Mini-prep
S05192-B0015 -
Antarctic phosphatase
S05192-B0015 -
Ligation
Ligate S05192-B0015 with R0040 -
Transformation
R0040-S05192-B0015
5µL DNA to DH5α -
Culture
R0040-S05192-B0015 -
Mini-prep
R0040-S05192-B0015 -
Complete!
