Exeter/9 July 2013

Our requested DNA has arrived from iGEM!

The Parts sent over were...

- Magenta pigment coding ([http://parts.igem.org/Part:BBa_K592012 BBa_K592012], pSB1C3)

- Green light sensor ([http://parts.igem.org/Part:BBa_K592001 BBa_K592001], pSB1C3)

- Yellow pigment coding ([http://parts.igem.org/Part:BBa_K592010 BBa_K592010], pSB1C3)

- Cyan pigment coding ([http://parts.igem.org/Part:BBa_K322122 BBa_K322122], pSB1C3)

- FixJ intermediate protein coding ([http://parts.igem.org/Part:BBa_K592005 BBa_K592005], pSB1C3)

We also requested some extra parts from the Registry

- 2 Genome Integration Kits ([http://parts.igem.org/Part:BBa_K510000 BBa_K510000] and [http://parts.igem.org/Part:BBa_K510012 BBa_K510012]). We know we want to genome integrate some of our Parts at some point...

- Alternative cyan pigments ([http://parts.igem.org/Part:BBa_K592011 BBa_K592011] just codes for the pigment but has no promoter, RBS or terminator. [http://parts.igem.org/Part:BBa_K864404 BBa_K864404] codes for the same pigment, but has a promoter and RBS. [http://parts.igem.org/Part:BBa_K592022 BBa_K592022] also has the same coding region and RBS, but an alternative promoter)

Afternoon

Transforming the BioBricks we will be utilising in the blue light module

We are transforming...

- [http://parts.igem.org/Part:BBa_K608002 BBa_K608002], codes for a promoter and RBS

- [http://parts.igem.org/Part:BBa_K592004 BBa_K592004], our blue light sensor

- [http://parts.igem.org/Part:BBa_B0015 BBa_B0015], a terminator

- [http://parts.igem.org/Part:BBa_B0034?title=Part:BBa_B0034 BBa_B0034], an RBS

- [http://parts.igem.org/Part:BBa_K592006 K592006], the promoter that is activated by FixJ

We already have the following Parts as LB stab plates from the iGEM registry, so a transformation of these Parts is required:

- [http://parts.igem.org/Part:BBa_K592005 BBa_K592005], codes for our intermediate protein, FixJ

- [http://parts.igem.org/Part:BBa_K592010 BBa_K592010], our yellow pigment

The transformation protocol from 4/7/13 was followed, no details from the method were changed.

Sequencing

We also sent our Cph8 plasmids off for sequencing.

Liquid cultures

Liquid cultures were made of...

- [http://parts.igem.org/Part:BBa_K592012 BBa_K592012], our magenta pigment

- [http://parts.igem.org/Part:BBa_K592010 BBa_K592010], our yellow pigment

- [http://parts.igem.org/Part:BBa_K322122 BBa_K322122], our cyan pigment

(Next day, results from transformation)

[http://parts.igem.org/Part:BBa_K592004 BBa_K592004], blue light sensor - 94 colonies

[http://parts.igem.org/Part:BBa_K608002 BBa_K608002], promoter and RBS - 112 colonies

[http://parts.igem.org/Part:BBa_K592006 BBa_K592006], FixJ promoter - 73 colonies

[http://parts.igem.org/Part:BBa_B0015 BBa_B0015], a terminator - 118 colonies

Our BBa_B0034, an RBS, had no colonies as we hadn't noted that this part was stored on pSB1A2, so plating it onto a chloramphenicol plate killed all of the colonies. A retry of this transformation will be undertaken tomorrow.

Take me back to the notebook.