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Exeter/9 July 2013
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'''Sequencing''' | '''Sequencing''' | ||
| - | We also sent our | + | We also sent our Cph8 plasmids off for sequencing. |
'''Liquid cultures''' | '''Liquid cultures''' | ||
Revision as of 10:42, 19 July 2013
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 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.
