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Team:Groningen/Notebook/2 September 2009
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(Difference between revisions)
m (New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== :→ {{todo}} Choose colonies from plates for growth of E.coli TOP10 with GVP behind LAC/pBAD (+RBS) promoter in pSB1AC3...) |
m (→Metal Accumulation) |
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===Metal Accumulation=== | ===Metal Accumulation=== | ||
| + | ====MBP-ArsR==== | ||
| + | *{{todo|Check sequence}} | ||
| + | *{{todo|Put promotors in front}} | ||
| + | *{{todo|Put transporters in back}} | ||
| + | ====fMT==== | ||
| + | *{{todo|Plasmid isolation & Restriction analysis}} | ||
| + | *{{todo|Put promotors in front}} | ||
| + | ====SmtA==== | ||
| + | *{{todo|Plasmid isolation & Restriction analysis}} | ||
| + | *{{todo|Put promotors in front}} | ||
===Vectors=== | ===Vectors=== | ||
Revision as of 07:39, 2 September 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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Wet
GVP Cluster
- → TODO Choose colonies from plates for growth of E.coli TOP10 with GVP behind LAC/pBAD (+RBS) promoter in pSB1AC3
- → TODO Choose colonies from plates for growth of E.coli TOP10 with [http://partsregistry.org/Part:pSB2K3 pSB2K3] plasmid from the registry (plate 1, 7C with RFP Coding Device [http://partsregistry.org/Part:BBa_J04450 BBa_J04450]
- → TODO Make glycerol stocks of pBad/araC, pBad/araC+RBS+GVP, and pLacI+RBS+GVP (both in pSB1A2 and pSB1AC3 plasmid)
- → TODO Test control of bouyancy in Saline solution (grow plates with GVP constructs)
- → TODO Order synthetic DNA for GVP
- → TODO Order primer for PstI site removal
- → TODO Test promoter strenght compared to BBa_J23101 promoter (Sven)
- → TODO Enter sequences of constructs to Sandbox
Colonies on Plates
| Name | Plasmid Used | Antibiotics on Plasmid | No. of Colonies | Date |
| [http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_J04450 BBa_J04450] (1,7C) | pSB2K3 | Kanamycin | 0 | 2/9 |
| [http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_J04450 BBa_J04450] (1,7C)(conc.) | pSB2K3 | Kanamycin | 4 | 2/9 |
| [http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_P1010 BBa_P1010] (1,7K) | pSB2K3 | Kanamycin | 0 | 2/9 |
| [http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_P1010 BBa_P1010] (1,7K)(conc.) | pSB2K3 | Kanamycin | 0 | 2/9 |
- → The plates showed very low colony growth, and can be caused by the fact it is a low copy plasmid which can be induced by iptg to increase the copy number. Additional growth with IPTG should increase the amount of colonies and available plasmid.
- → From the plate with four colonies 5mL LB-amp100 medium was inoculated, and additional plates were swiped with the used colonies to see if single colony growth on plate can occur.
Transporters
Metal Accumulation
MBP-ArsR
- Check sequence
- Put promotors in front
- Put transporters in back
fMT
- Plasmid isolation & Restriction analysis
- Put promotors in front
SmtA
- Plasmid isolation & Restriction analysis
- Put promotors in front
Vectors
Dry
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