"
Exeter/16 August 2013
From 2013.igem.org
Adam + Tom
Miniprep of our complete Cph8 for inertion into CAM plasmid to be sent into the registry.
Miniprep of Complete Cph8 and Phycocyanobilin Operon (part [http://parts.igem.org/Part:BBa_K322122 BBa_K322122])
Realised we had ran out of CAM plasmid, therefore had to transform some more out of the kit plate.
Made transformations of:
TET plasmid
CAM plasmid
KAN plasmid
AMP plasmid
from the kit plates.
Checking gBlocks of Green Light module
We have three plates of our gBlocks coding for the green light module, Two show good growth, the third appears to have no growth whatsoever. Liquid cultures of all three plates were made on 15/8/13, with 16 colonies being selected from each of the three plates (48 liquid cultures in all). All liquid cultures for plate 3 (no visible cultures) had zero growth (as expected) but the liquid cultures for plates one and two were successful.
These were Miniprepped this morning, and then run on gels after being cut with EcoRI and PstI. The gels are shown below.
Gel for Plate 1, colonies 1 through 10
Run against 1kb GeneRuler.
Strangely, the ladder in lane 1 appears to have not worked, but the ladder in lane 12 (far RHS) has. This suggests there may be something wrong with the gel, which is supported by the fact that we can't see any bands for any colonies.
Gel for Plate 1, colonies 11 through 16
Run against 1kb GeneRuler.
The gel appears to have worked. Each colony shows the same bands at the same sizes, which is promising. The band at ~2,000bp should be the plasmid, the band above it near 3,000bp should be the gBlocks making up the green light module.
Gel for Plate 1, colonies 11 through 16
Run against 1kb GeneRuler.
.jpg)
This gel is not as successful as hoped. The second half is missing (human error, I'm afraid!) but showed that both ladders had worked, but no strong DNA bands were present in the main portion of the gel. There are very faint bands, but they are highly smudged and indistinguishable from the background noise.
Take me back to the notebook.
With many thanks to our sponsors:
