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Team:HokkaidoU Japan/Notebook/plastic Week 13
From 2012.igem.org
(Difference between revisions)
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We got colonies on the plate, and started liquid culture. | We got colonies on the plate, and started liquid culture. | ||
| - | ==PCR== | + | ====PCR==== |
<p> | <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> | </p> | ||
| - | ==Digestion== | + | ====Digestion==== |
<p> | <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. | ||
</p> | </p> | ||
| - | ==Ligation== | + | ====Ligation==== |
<p> | <p> | ||
Ligated the promoter with RBS-PhaC-RBS-PhaA-RBS-PhaB-dT and with pSB1A2. | Ligated the promoter with RBS-PhaC-RBS-PhaA-RBS-PhaB-dT and with pSB1A2. | ||
</p> | </p> | ||
| - | ==Transformation== | + | ====Transformation==== |
<p> | <p> | ||
Transformed each ligation product into JM109, and incubated. | Transformed each ligation product into JM109, and incubated. | ||
Revision as of 22:05, 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
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.
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.
