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Team:HokkaidoU Japan/Notebook/Color R0040-B0034-(K592010, K592012, K864401)-B0015
From 2013.igem.org
Maestro E.coli
Notebook
R0040-B0034-(K592010, K592012, K864401)-B0015
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Start
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Get chromoprotein DNA
Great thanks to iGEM Uppsala! -
Transformation
K592010, K592012, K864401
each of 1µL DNA to DH5α -
Colony PCR & Culture
K592010, K592012, K864401, B0015
We confirmed transformation was completely successful.
Cultured them for subsequent experiments. -
Mini-prep
K592010, K592012, K864401, B0015 -
PCR & PCR purification
K592010, K592012, K864401
100b up, 200b dn, KOD Plus NEO -
Digestion
Cut B0015 with EcoRI, XbaI (using 10xM buffer)
Cut K592010, K592012, K864401 with EcoRI, SpeI (using 10xH buffer) -
Antarctic phosphatase
B0015 -
Ligation
Ligate B0015 with K592010, K592012, K864401 -
Transformation
(K592010, K592012, K864401)-B0015
each of 5µL DNA to DH5α -
PCR & PCR purification
R0040-B0034 from our stock
100b up, 200b dn, KOD Plus NEO -
Colony PCR
(K592010, K592012, K864401)-B0015 -
Culture
(K592010, K592012, K864401)-B0015 -
Mini-prep
(K592010, K592012, K864401)-B0015 -
Digestion
Cut K592010-B0015 with EcoRI, XbaI (using 10xM buffer)
Cut R0040-B0034 with EcoRI, SpeI (using 10xH buffer) -
Antarctic phophatase
(K592010, K592012, K864401)-B0015 -
Ligation
Ligate (K592010, K592012, K864401)-B0015 with R0040-B0034 -
Transformation
R0040-B0034-(K592010, K592012, K864401)-B0015
each of 5µL DNA to DH5α -
Culture
R0040-B0034-(K592010, K592012, K864401)-B0015 -
Mini-prep
R0040-B0034-(K592010, K592012, K864401)-B0015 -
Complete!
