Exeter/9 July 2013

From 2013.igem.org

Exeter iGEM 2013 · Paint by Coli

Our requested DNA has arrived from iGEM!

The Parts sent over were...

- Magenta pigment coding (BBa_K592012, pSB1C3)

- Green light sensor (BBa_K592001, pSB1C3)

- Yellow pigment coding (BBa_K592010, pSB1C3)

- Cyan pigment coding (BBa_K322122, pSB1C3)

- FixJ intermediate protein coding (BBa_K592005, pSB1C3)

We also requested some extra parts from the Registry

- 2 Genome Integration Kits (BBa_K510000 and BBa_K510012). We know we want to genome integrate some of our Parts at some point...

- Alternative cyan pigments (BBa_K592011 just codes for the pigment but has no promoter, RBS or terminator. BBa_K864404 codes for the same pigment, but has a promoter and RBS. 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...

- BBa_K608002, codes for a promoter and RBS

- BBa_K592004, our blue light sensor

- BBa_B0015, a terminator

- BBa_B0034, an RBS

- 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:

- BBa_K592005, codes for our intermediate protein, FixJ

- 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...

- BBa_K592012, our magenta pigment

- BBa_K592010, our yellow pigment

- BBa_K322122, our cyan pigment

(Next day, results from transformation)

BBa_K592004, blue light sensor - 94 colonies

BBa_K608002, promoter and RBS - 112 colonies

BBa_K592006, FixJ promoter - 73 colonies

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.

Exeter iGEM 2013 · Paint by Coli