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Team:Groningen/Notebook/30 August 2009
From 2009.igem.org
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|[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | |[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | ||
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|[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | |[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | ||
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|[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | |[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | ||
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|[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | |[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | ||
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|[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | |[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 <i>E. coli</i> TOP10] | ||
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==Transformation== | ==Transformation== | ||
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Revision as of 08:55, 30 August 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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Weekend stuff
Metal accumulation
| Growth | Name | Plasmid | Inoculation date | Antibiotics |
|---|---|---|---|---|
| pBAD | pSB1AC3? | 29august2009 | LB-Cam | |
| pBAD | pSB1AC3? | 29august2009 | LB-Amp | |
| pLac | pSB1AC3? | 29august2009 | LB-Cam | |
| pLac | pSB1AC3? | 29august2009 | LB-Amp | |
| fMT #6 | pSB1AC3 | 29august2009 | LB-Amp | |
| SmtA #3 | pSB1AC3 | 29august2009 | LB-Amp | |
| pLow-fMT #3 | pSB1AC3 | 29august2009 | LB-Amp | |
| pLow-fMT #4 | pSB1AC3 | 29august2009 | LB-Amp | |
| pLac-fMT #2 | pSB1AC3 | 29august2009 | LB-Amp |
pSB1AC3 There was doubt on the identity of the vector in which the inducible promotors were build, seeing as it did not grow on LB supplemented with chloramphenicol it is most probably not pSB1AC3, perhaps pSB1A2. Cultures were put in glycerol stocks, except for pBAD & pLac whose identity first need to be confirmed.
Glycerol stocks (-80 °C)
| Component | Description | Part or Accession # | Base Pairs (bp) | Plasmid (backbone) | Resistance | Well | Quality control | Storage Medium+Antibiotics | Organism | Date of storage |
| pLac-fMT | Metallothionein with inducible promotor | Part or Accession # | Base Pairs (bp) | pSB1AC3 | Ampicillin/chloramphenicol | Well | Quality control | LB-Ampicillin | [http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 E. coli TOP10] | 30august2009 |
| pLow-fMT #4 | Metallothionein with low constitutive promotor | Part or Accession # | Base Pairs (bp) | pSB1AC3 | Ampicillin/chloramphenicol | Well | Quality control | LB-Ampicillin | [http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 E. coli TOP10] | 30august2009 |
| pLow-fMT #3 | Metallothionein with low constitutive promotor | Part or Accession # | Base Pairs (bp) | pSB1AC3 | Ampicillin/chloramphenicol | Well | Quality control | LB-Ampicillin | [http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 E. coli TOP10] | 30august2009 |
| SmtA | Metallothionein | Part or Accession # | Base Pairs (bp) | pSB1AC3 | Ampicillin/chloramphenicol | Well | Quality control | LB-Ampicillin | [http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 E. coli TOP10] | 30august2009 |
| fMT #6 | Metallothionein | Part or Accession # | Base Pairs (bp) | pSB1AC3 | Ampicillin/chloramphenicol | Well | Quality control | LB-Ampicillin | [http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29 E. coli TOP10] | 30august2009 |
Transformation
| Colonies | Name | Plasmid | Date of plating | Antibiotics |
|---|---|---|---|---|
| Negative control (50 μL) | 1 μL MilliQ | 29august2009 | LB-Amp100 | |
| Negative control (200 μL) | 1 μL MilliQ | 29august2009 | LB-Amp100 | |
| Positive control (50 μL) | 1 μL pSB1AC3+high constitutive promotor | 29august2009 | LB-Amp100 | |
| Positive control (200 μL) | 1 μL pSB1AC3+high constitutive promotor | 29august2009 | LB-Amp100 | |
| SmtA+pLac (50 μL) | pSB1AC3 | 29august2009 | LB-Amp100 | |
| SmtA+pLac (200 μL) | pSB1AC3 | 29august2009 | LB-Amp100 | |
| SmtA+pLow (50 μL) | pSB1AC3 | 29august2009 | LB-Amp100 | |
| SmtA+pLow (200 μL) | pSB1AC3 | 29august2009 | LB-Amp100 |
pSB1AC3 was in fact pSB3K3! Therefore no colonies could grow.
5 colonies of each promotor were inoculated for an ON culture, colony PCR was postponed to monday.
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