Exeter/23 July 2013
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Revision as of 11:25, 25 July 2013 by Fentwistle (Talk | contribs)
Unfortunately, some of our liquid cultures from yesterday appear to have not worked.
Also, the liquid cultures for OmpR and the OmpR promoter were mislabelled, so we're uncertain which is which. However, when we run "OmpR A" on a gel, it should be very obvious which is which, as OmpR is 917bp and the OmpR promoter is 108bp.
Part | Did it work? | Source |
---|---|---|
NP1 (CcaS + Terminator) | ✗ | Transformation plate |
NP2 (promoter, RBS, FixJ + Terminator | ✗ | Transformation plate |
NP3 (yellow pigment + Terminator) | ✗ | Transformation plate |
NP4 (magenta pigment + Terminator) | ✗ | Transformation plate |
NP7 (YF1 + Terminator) | ✓ | Transformation plate |
NP8 (FixJ + Terminator) | ✓ | Transformation plate |
Ligation control | ✗ | Transformation plate |
Lambda inverter system | ✗ | Streak plate in fridge |
T7 promoter | ✗ | Streak plate in fridge |
Promoter + RBS (BBa_K608002) | ✓ | Streak plate in fridge |
RBS (BBa_B0034) | ✓ | Streak plate in fridge |
OmpR "A" | ✓ | Streak plate in fridge |
OmpR "B" | ✗ | Streak plate in fridge |
RFP control | ✓ | Transformation plate |
RBS + cph8 (BBa_K592018) | ✓ | Transformation plate |
The working liquid cultures (NP7, NP8, Promoter + RBS, RBS, OmpR "A" and RBS + cph8) were MiniPrepped, then NanoDropped, then had their concentrations increased using the SureClean protocol from 17/7/13. Each culture was replicated twice.
Part | NanoDrop data (ng/ul) | NanoDrop data after SureClean protocol (ng/ul) |
---|---|---|
Promoter and RBS #1 | 3.5 | 107.3 |
Promoter and RBS #2 | 2.1 | 107.3 |
NP8 #1 | 4.5 | 113.3 |
NP8 #2 | 5.9 | 40.1 |
RBS (BBa_B0034) #1 | 14.0 | 64.8 |
RBS (BBa_B0034) #2 | 2.2 | Would not pellet, SureClean failed |
NP7 #1 | 35.5 | SureClean not required |
NP7 #2 | 29.3 | SureClean not required |
Ompr "A" #1 | 2.1 | 110.5 |
OmpR "A" #2 | 2.0 | 19.5 |
RBS and cph8 #1 | 3.5 | 197.1 |
RBS and cph8 #2 | 12.5 | Would not pellet, SureClean failed |