Team:HokkaidoU Japan/Notebook/plastic Week 12

From 2012.igem.org

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==September 18th==
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==Colony PCR==
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We confirmed the succession of ligation RBS-PhaC-RBS-PhaA-RBS-PhaB-dT with pTet on pSB1A2 by colony PCR.<br/>
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RBS-PhaB-dT, a part of insert was multiplied.
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==Secuencing==
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The sequence of RBS-PhaC-RBS-PhaA-RBS-PhaB-dT was analyzed.
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The result showed that...
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Revision as of 03:15, 18 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.

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...