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Team:HokkaidoU Japan/Notebook/plastic Week 12
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| + | <div class="hokkaidou-notebook-daily"> | ||
| + | ==September 17th== | ||
| + | <div> | ||
| + | ==Plasmid extraction== | ||
| + | <p> | ||
| + | Plasmids of RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2 and pTet(BBa_0040) on pSB1A2 were extracted.<br/> | ||
| + | And then we got DNA solution of them. | ||
| + | </p> | ||
| + | |||
| + | ==Digestion== | ||
| + | <p> | ||
| + | RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2 was digested with XbaI and PstI restriction sites.<br/> | ||
| + | pTet on pSB1A2 was also digested with SpeI and PstI. | ||
| + | </p> | ||
| + | |||
| + | ==Gel extraction== | ||
| + | <p> | ||
| + | We confirmed the succession of digestion by electrophoresis.<br/> | ||
| + | And then DNA were extracted from TBE gel. | ||
| + | </p> | ||
| + | |||
| + | ==Ethanol precipitation== | ||
| + | <p> | ||
| + | The digested DNAs were condensed by Ethanol precipitation. | ||
| + | </p> | ||
| + | |||
| + | ==Ligation== | ||
| + | <p> | ||
| + | RBS-PhaC-RBS-PhaA-RBS-PhaB-dT was ligated with pTet on pSB1A2. | ||
| + | </p> | ||
| + | |||
| + | ==Transformation== | ||
| + | <p> | ||
| + | The ligated DNA was transformed into E.coli (strain: JM109).<br/> | ||
| + | And then we spread fungus liquid on LBA plates. | ||
| + | </p> | ||
| + | |||
| + | </div></div> | ||
<!-- DO NOT EDIT UNDER THIS LINE @iTakeshi --> | <!-- DO NOT EDIT UNDER THIS LINE @iTakeshi --> | ||
Revision as of 13:04, 17 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.
