iGEM 2009 — Team SDU-Denmark

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# Team:SDU-Denmark/Notebook

### From 2009.igem.org

| [Home](/Team%3ASDU-Denmark "Team:SDU-Denmark") | [The Team](/Team%3ASDU-Denmark/Team "Team:SDU-Denmark/Team") | [The Project](/Team%3ASDU-Denmark/Project "Team:SDU-Denmark/Project") | [Parts Submitted to the Registry](/Team%3ASDU-Denmark/Parts "Team:SDU-Denmark/Parts") | [Modeling](/Team%3ASDU-Denmark/Modeling "Team:SDU-Denmark/Modeling") | **Notebook** |
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| Contents   * [1 Project task-management notebook](#Project_task-management_notebook)   + [1.1 Todo](#Todo)     - [1.1.1 Create biobrick RIP in registry](#Create_biobrick_RIP_in_registry)     - [1.1.2 Create biobrick RIP+Export in registry](#Create_biobrick_RIP.2BExport_in_registry)   + [1.2 Doing](#Doing)   + [1.3 Done](#Done)     - [1.3.1 Find Plasmid backbone](#Find_Plasmid_backbone)     - [1.3.2 E. coli strain](#E._coli_strain)     - [1.3.3 Promoter for e.coli](#Promoter_for_e.coli)     - [1.3.4 RBS](#RBS)     - [1.3.5 Terminator](#Terminator)   + [1.4 Done Done](#Done_Done) |

# Project task-management notebook

## Todo

### Create biobrick RIP in registry

### Create biobrick RIP+Export in registry

## Doing

## Done

### Find Plasmid backbone

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

**People:** Helle and Mike

*July 7th 2009:*
We found the backbone, [http://partsregistry.org/wiki/index.php/Part:BBa\_I742123 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 [http://partsregistry.org/Part:pSB1A3 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)

### E. coli strain

**Task:** Find a e.coli strain

**People:** Mike

*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 first.

### Promoter for e.coli

**Task:** Find a constitutive and inducible promoter

**People:** Anna and Kir

*July 7th 2009*

**Constitutive promoter**

We'll use [http://partsregistry.org/Part:BBa\_J23100 BBa\_J23100] as constitutive promoter. Its located in QC09 Kit Plate 1, Well C18.

**Inducible promoter**

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

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.

### RBS

**Task:** Find a RBS

**People:** John and Julius

*July 7th 2009*

We'll use [http://partsregistry.org/wiki/index.php?title=Part:BBa\_B0034 BBa\_B0034]. The sequence is: 5'-AAAGAGGAGAAA-3'. It's located in: Kit plate 1, well 2M.

Binding to the site is not regulated, and it has been reported to be very effective.

### Terminator

**Task:** Find a terminator

**People:** Marc

*July 7th 2009*

We'll use [http://partsregistry.org/wiki/index.php?title=Part:BBa\_B0015 BBa\_B0015]. This is a double terminator consisting of BBa\_B0010 and BBa\_B0012. This is the most commonly used terminator. It seems to be reliable.
Note, however, that BBa\_B0014 is a better part for forward and reverse termination.

It's located in: QC09 Plate 669, Well 41811.

## Done Done

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