IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/06/13/Min Lin

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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.

Check up parts in partsregistry.

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

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 )

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