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.

15:00
Check up parts in partsregistry.
Results:

BBa_E0240Rbs+GFP+terminatorMedium rbs1-12MAmp
BBa_E0840Rbs+GFP+terminatorStrong rbs1-12OAmp
BBa_R0080Ara-Promoter 1-12EAmp
BBa_C0062LuxR geneNo LVA1-4OAmp
BBa_C0051CI repressor+LVA1-4EAmp
BBa_C0179LasR activatorNo LVA2-8MAmp
BBa_C0079LasR activatorLVA1-14JKan
BBa_R0079LasR/PAI promoter 1-12AAmp

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

EcoRI0.5μL
SpeI0.5μL
10xH buffer2μL
Plasmid 10μL
ddH2O7μL
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