Team:HokkaidoU Japan/Notebook/plastic Week 12


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September 17th

Plasmid extraction

Plasmids of RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2 and pTet(BBa_0040) on pSB1A2 were extracted.
And then we got DNA solution of them.


RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2 was digested with XbaI and PstI restriction sites.
pTet on pSB1A2 was also digested with SpeI and PstI.

Gel extraction

We confirmed the succession of digestion by electrophoresis.
And then DNA were extracted from TBE gel.

Ethanol precipitation

The digested DNAs were condensed by Ethanol precipitation.


RBS-PhaC-RBS-PhaA-RBS-PhaB-dT was ligated with pTet on pSB1A2.


The ligated DNA was transformed into E.coli (strain: JM109).
And then we spread fungus liquid on LBA plates.

Colony PCR

We confirmed the ligation of [RBS-B] and pSB1C3. The results showed that the ligation went well. We chose three colonies and started the liquid culture.

liquid culture

The colony of [RBS-phaC-RBS-phaA on pSB1A2] and [RBS-phaC-RBS-phaA-RBS-phaC-dT on pSB1A2] was condensed in LB.

September 18th

Colony PCR

We confirmed the succession of ligation RBS-PhaC-RBS-PhaA-RBS-PhaB-dT with pTet on pSB1A2 by colony PCR.
RBS-PhaB-dT, a part of insert was multiplied.


The sequence of RBS-PhaC-RBS-PhaA-RBS-PhaB-dT was analyzed. The result showed that...

Plasmid extraction

The plasmids, [RBS-PhaC-RBS-PhaA] on pSB1A2, [RBS-PhaB] on pSB1C3 and [RBS-PhaC-RBS-PhaA-RBS-PhaB-dT] on pSB1A2 were extracted.