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Team:Groningen/Notebook/17 August 2009
From 2009.igem.org
(Difference between revisions)
(New page: {{Team:Groningen/Notebook/Day/Header}} ==Wet== ===GVP Cluster=== :→ {{todo}} Restriction for Assembly :→ {{todo}} Gel purification of plasmid :→ {{todo}} Ligation of metal promote...) |
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:→ {{todo}} Transformation of E.coli TOP10 cells | :→ {{todo}} Transformation of E.coli TOP10 cells | ||
:→ {{todo}} Test promoter/GVP constructs (grow precultures) | :→ {{todo}} Test promoter/GVP constructs (grow precultures) | ||
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'''Tranformation''' | '''Tranformation''' | ||
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===Transporters=== | ===Transporters=== | ||
Revision as of 07:05, 17 August 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 Restriction for Assembly
- → TODO Gel purification of plasmid
- → TODO Ligation of metal promoter oligo's into vector BBa_J61002
- → TODO Transformation of E.coli TOP10 cells
- → TODO Test promoter/GVP constructs (grow precultures)
- → TODO Grow o.n. cultures to check ligation of pSB3K3-H/L-GVP constructs of last week
Restriction for Assembly
The vector [http://partsregistry.org/wiki/index.php?title=Part:BBa_J61002 BBa_J61002] containing the [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 high] constitutive promoter was cut with EcoRI and SpeI to create correct ends for insert of metal promoter oligo's.
- 8 μL plasmid in MQ (1.0μg)
- 8 μL MQ (end volume of 20μL)
- 2μL Fast digest buffer
- 1μL PstI fast digest enzyme
- 1μL SpeI fast digest enzyme
Restriction was kept at 37C for 30 min. and put on ice until used for gel purification.
Purification
Concentrations
| Plasmid | Conc. ng/μL | 260/280 | 260/230 | -20 box (michael | Restriction Control |
| BBa_J61002-H SpeI/EcoRI | ? | ? | ? | ? | ? |
Ligation
Tranformation
Transporters
Metal Accumulation
Vectors
Dry
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