IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/06/13/Min Lin
13:00 pick one colony of Top10 containing T7 promoter plasmid. Incubate in 5 ml LB (Kan+).
14:00 adjust DNA imager with a mixture of 20ul Marker and 5ul genefinder. Image successfully captured.
13:00~14:00 transformation
Transform Plasmid pAra-T7ptag from Voigt lab to Top10 competent.
0.5ul plasmid diluted into 1ul. Then transform.
Add 500ul LB without antibiotic and shake in the incubator for 40 minutes.
Centrifuge to concentrate E.coli then Plate.
Keep the plate in 37 centigrade incubator.
15:00
Check up parts in partsregistry.
Results:
| BBa_E0240 | Rbs+GFP+terminator | Medium rbs | 1-12M | Amp |
| BBa_E0840 | Rbs+GFP+terminator | Strong rbs | 1-12O | Amp |
| BBa_R0080 | Ara-Promoter | 1-12E | Amp | |
| BBa_C0062 | LuxR gene | No LVA | 1-4O | Amp |
| BBa_C0051 | CI repressor | +LVA | 1-4E | Amp |
| BBa_C0179 | LasR activator | No LVA | 2-8M | Amp |
| BBa_C0079 | LasR activator | LVA | 1-14J | Kan |
| BBa_R0079 | LasR/PAI promoter | 1-12A | Amp |
Dissolve parts in 15ul ddH2O store in PCR tube in -20
15:00
Sterile LB.
For plates(300ml+300ul Amp)
Liquid LB: 300ml with no antibiotics. 300ml with Amp. Dry in the hood.
21:00 Transformation ( BBa_E0240&BBa_E0840 Top10 )
23:00
MiniPrep T7 promoter plasmid.
Digest with EcoRI and SpeI 37 centigrade over night
| EcoRI | 0.5μL |
| SpeI | 0.5μL |
| 10xH buffer | 2μL |
| Plasmid | 10μL |
| ddH2O | 7μL |
