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Team:HokkaidoU Japan/Notebook/RBS SD8-1C3-Replacement
From 2013.igem.org
Maestro E.coli
Notebook
SD8-1C3 Replacement
-
Start
-
Mini-Prep
oripro on 1C3 -
PCR
R0040-SD8-LacZα-B0015 100up 200dn -
Gel Extraction
R0040-SD8-LacZα-B0015 -
Digestion
Cut R0040-SD8-LacZα-B0015 on 1A2 and Oripro-B0034-LacZα-B0015 on 1C3 EcoRI and PstI (using 10×H buffer) -
Gel Extraction
Oripro-B0034-LacZα-B0015 R0040-SD8-LacZα-B0015 -
Ethanol Precipitation
Oripro-B0034-LacZα-B0015 R0040-SD8-LacZα-B0015 -
Ligation
Ligate R0040-SD8-LacZα-B0015 with Vector(pSB1C3) -
Transformation
R0040-SD8-LacZα-B0015
each of 5µL DNA to DH5α
faild -
Transformation
R0040-SD8-LacZα-B0015
each of 5µL DNA to DH5α -
Colony PCR
R0040-SD8-LacZα-B0015 -
Culture
R0040-SD8-LacZα-B0015 -
Mini-prep
R0040-SD8-LacZα-B0015 -
Complete!
