Team:SDU-Denmark/Notebook

From 2009.igem.org

(Difference between revisions)
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'''Task:''' Find a backbone for both e.coli and b. subtilis
'''Task:''' Find a backbone for both e.coli and b. subtilis
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'''Status:''' Doing.
 
'''People:''' Helle and Mike
'''People:''' Helle and Mike
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'''Last edit:''' July 8th 2009.
 
''July 7th 2009:''  
''July 7th 2009:''  
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'''Task:''' Find a promoter, that can be induced
'''Task:''' Find a promoter, that can be induced
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'''Status:''' Doing.
 
'''People:''' Anna
'''People:''' Anna
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'''Last edit:''' July 8th 2009.
 
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'''Task:''' Find a e.coli strain
'''Task:''' Find a e.coli strain
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'''Status:''' Doing.
 
'''People:'''  
'''People:'''  
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'''Last edit:''' July 8th 2009.
 
''July 7th 2009''
''July 7th 2009''
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The plasmid seems to be working in most types (no need for ccdB)
The plasmid seems to be working in most types (no need for ccdB)
Jacob suggests E. coli KG22, which can turns on the operon by adding IPTG.
Jacob suggests E. coli KG22, which can turns on the operon by adding IPTG.
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''July 8th 2009''
''July 8th 2009''
We'll use Top10 e.coli strain
We'll use Top10 e.coli strain
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===Promoter for e.coli===
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'''Task:''' Find a constitutive and inducible promoter
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'''People:'''
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''July 7th 2009''
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''''Constitutive promoter''''
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We'll use [http://partsregistry.org/Part:BBa_J23100 BBa_J23100] as constitutive promoter. Its located in QC09 Kit Plate 1, Well C18.
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''''Inducible promoter''''
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We'll use [http://partsregistry.org/Part:BBa_R0011 BBa_R0011] as an inducible promoter. It's located in: QC09 Kit Plate 1, Well 6G, Plasmid: pSB1A2
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Use:
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To induce this promoter region, generally add 1mM of IPTG to a plate (amount will vary by cell strain). That is, total volume of stuff in plate (usually around 0.025 liters) * 0.001 = X amount of liters that you will add to the solution.
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Revision as of 08:58, 9 July 2009

Home The Team The Project Parts Submitted to the Registry Modeling Notebook


Contents

Project task-management notebook

Todo

Create biobrick RIP in registry

Create biobrick RIP+Export in registry

Doing

Plasmid backbone

Task: Find a backbone for both e.coli and b. subtilis

People: Helle and Mike


July 7th 2009: We found the backbone, BBa_I742123, which other teams have found to be compatible with both both gram positive and gram negative bacteria, such as e. coli.

After talking to HQ, it turns out that the quality of their stock is bad, and they will try to get some home from older teams. Furthermore, it might simply be better to use a more specifik backbone for e. coli and for b. subtilis.

(mike)


July 8th 2009: We decided to start working with pSB1A3 instead, which is a high-output e. coli backbone.

Output: 100-300 number pr. cell. Resistance: ampicillin (we're going with 50 µg/mL first)

(mike)

Inducible promoter

Task: Find a promoter, that can be induced

People: Anna


July 7th 2009

I found the inducible promoter BBa_R0011 that is regulated by LacI.

It is a strong promoter, that will be on in stains without LacI.

(anna)


E. coli strain

Task: Find a e.coli strain

People:


July 7th 2009

The plasmid seems to be working in most types (no need for ccdB) Jacob suggests E. coli KG22, which can turns on the operon by adding IPTG.


July 8th 2009

We'll use Top10 e.coli strain


Promoter for e.coli

Task: Find a constitutive and inducible promoter

People:


July 7th 2009


'Constitutive promoter'

We'll use BBa_J23100 as constitutive promoter. Its located in QC09 Kit Plate 1, Well C18.


'Inducible promoter'

We'll use BBa_R0011 as an inducible promoter. It's located in: QC09 Kit Plate 1, Well 6G, Plasmid: pSB1A2

Use:

To induce this promoter region, generally add 1mM of IPTG to a plate (amount will vary by cell strain). That is, total volume of stuff in plate (usually around 0.025 liters) * 0.001 = X amount of liters that you will add to the solution.


Done

Done Done