Team:HokkaidoU Japan/Notebook/RBS SD2,4,6,8-3K3-Replacement
From 2013.igem.org
Maestro E.coli
Notebook
SD2, 4, 6, 8-3K3 Replacement
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Start
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PCR
R-0040-SD(2,4,6,8)-LacZα-B0015 100up 200dn -
Digestion
R-0040-SD(2,4,6,8)-LacZα-B0015 EcoRI and PstI (uning 10×H buffer)
pSB3K3 vector EcoRI and PstI (using 10×H buffer) -
Antarctic phophatase
pSB3K3 vector -
Ligation
R-0040-SD(2,4,6,8)-LacZα-B0015 with pSB3K3 -
Transformation
R0040-SD(6/4/2)-LacZα-B0015
each of 1µL DNA to DH5α -
Mini-prep
R-0040-SD(2,4,6,8)-LacZα-B0015 on pSB3K3 -
Sequence
R-0040-SD(2,4,6,8)-LacZα-B0015 on pSB3K3 -
β-galactosidase assay (1)
R0040-RBS(SD2/4/6/8)-LacZα-B0015, X/S (as N. C.) -
β-galactosidase assay (2)
R0040-RBS(B0034/SD8)-LacZα-B0015, X/S (as N. C.) -
β-galactosidase assay (3)
R0040-RBS(B0034/SD8)-LacZα-B0015, X/S (as N. C.) -
Complete!