Team:HokkaidoU Japan/Notebook/plastic Week 13



September 24th

Single colony isolation

We got few colonies from the plate. The colonies were isolated to another plate.

September 25th

Liquid culture

We got colonies on the plate, and started liquid culture.


Did PCR for promoter of R.eutropha from pGEM to ligate with RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2.


Digested the promoter done PCR and RBS-PhaA on pSB1A2 with XbaI and SpeI, and plasmid of RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2 with XbaI.

Gel extraction

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

Ethanol precipitation

The digested DNAs were condensed by Ethanol precipitation.


Ligated the promoter with RBS-PhaC-RBS-PhaA-RBS-PhaB-dT and with pSB1A2.


Transformed each ligation product into JM109, and incubated.

September 26th

Colony PCR

Colony PCR for colonies transformed at 25th.