Team:HokkaidoU Japan/Notebook/plastic Week 13

From 2012.igem.org

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====PCR====
====PCR====
-
<p>
 
Did PCR for promoter of R.eutropha from pGEM to ligate with RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2.
Did PCR for promoter of R.eutropha from pGEM to ligate with RBS-PhaC-RBS-PhaA-RBS-PhaB-dT on pSB1A2.
-
</p>
 
====Digestion====
====Digestion====
-
<p>
 
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.
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.
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</p>
 
====Gel extraction====
====Gel extraction====
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<p>
 
We confirmed succession of digestion by electrophoresis.
We confirmed succession of digestion by electrophoresis.
And then DNA were extracted from TBE gel.
And then DNA were extracted from TBE gel.
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</p>
 
====Ligation====
====Ligation====
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====Transformation====
====Transformation====
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<p>
 
Transformed each ligation product into JM109, and incubated.
Transformed each ligation product into JM109, and incubated.
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</p>
 
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====Colony PCR====
====Colony PCR====
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<p>
 
Colony PCR for colonies transformed at 25th.
Colony PCR for colonies transformed at 25th.
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</p>
 
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</div></div>
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Revision as of 22:13, 26 September 2012

Contents

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.

PCR

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

Digestion

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.

Ligation

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

Transformation

Transformed each ligation product into JM109, and incubated.

September 26th

Colony PCR

Colony PCR for colonies transformed at 25th.


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