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Team:HokkaidoU Japan/Notebook/RBS SD2,4,6
From 2013.igem.org
Maestro E.coli
Notebook
SD2, 4, 6
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Start
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PCR
LacZα-B0015 100up 200dn -
PCR
R0040-SD8-LacZα-B0015 / SD8 100up SD(2/4/6) mutation revers -
PCR purification
R0040-SD(6/4/2) / SD(6/4/2) -
Digestion
Cut LacZα-B-0015 EcoRI and XbaI (using 10×M buffer) , PCR products EcoRI and SpeI (using 10×H buffer) -
Ethanol precipitation
R0040-SD(6/4/2) / SD(6/4/2)
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Gel extraction
LacZα-B-0015 -
Ethanol precipitation
LacZα-B-0015 -
Ligation
Ligate R0040-SD(6/4/2) / SD(6/4/2) with LacZα-B0015
Used X-cut-vector for inserts with R0040, and S-cut-vector for insrts without R0040 -
Transformation
R0040-SD(6/4/2)LacZα-B0015 / SD(6/4/2)
each of 4µL DNA to DH5α -
Sequence
SD(6/4/2) on pSB1C3 -
Complete!
