"
Team:HokkaidoU Japan/Notebook/plastic Week 12
From 2012.igem.org
| Line 38: | Line 38: | ||
The ligated DNA was transformed into E.coli (strain: JM109).<br/> | The ligated DNA was transformed into E.coli (strain: JM109).<br/> | ||
And then we spread fungus liquid on LBA plates. | And then we spread fungus liquid on LBA plates. | ||
| + | |||
</p> | </p> | ||
| + | |||
| + | ------- | ||
| + | |||
| + | ==Colony PCR== | ||
| + | |||
| + | <p> | ||
| + | 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. | ||
| + | </p> | ||
| + | |||
</div></div> | </div></div> | ||
| Line 56: | Line 69: | ||
The result showed that... | The result showed that... | ||
</p> | </p> | ||
| + | |||
| + | |||
| + | |||
<!-- DO NOT EDIT UNDER THIS LINE @iTakeshi --> | <!-- DO NOT EDIT UNDER THIS LINE @iTakeshi --> | ||
Revision as of 12:51, 22 September 2012
Contents |
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.
Digestion
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.
Ligation
RBS-PhaC-RBS-PhaA-RBS-PhaB-dT was ligated with pTet on pSB1A2.
Transformation
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.
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.
Secuencing
The sequence of RBS-PhaC-RBS-PhaA-RBS-PhaB-dT was analyzed. The result showed that...
