Team:Paris Liliane Bettencourt/Notebook/2010/07/21/
From 2010.igem.org
Antoine
Overnight culture
E.coli TG1 carrying pBM43 (mccABCDE+AmpR+CmR. 2 plates with two different kind of colonies both (big and small). Overnigth culture LB+Cm+Amp for:
- one big colony plate 1
- one small colony plate 1
- one big colony plate 2
- one small colony plate 2
Transformation Biobrick in TOP10
- PlTetO BBa_R0040
- TetR BBa_C0040
- PLacI BBa_R0011
- LacI BBa_C0012
All on Amp plate
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Théotime and Raphaël
Minipreps
2 minipreps for each biobrick from 5ml culture each. Elution of each miniprep with 50µl of Elution Buffer, which gives about 90µl of miniprep per biobrick.
- J732006 (LacZ) -exception : only 3 ml of culture left to work with
- B0031 (weak RBS)
- B0032 (medium RBS)
- B0034 (standard RBS)
- J31007 (TetA C)
- C0062 (LuxR w/o RBS)
- J37033 (RBS/LuxR) from the 2007 distribution plate
- 23119 (Ps) - Amp
- J23110 (Pm) - Amp
- attC -Kan
- term/attC/mRFP -Kan
- B0015 (term) - Amp
- E0240 (RBS/GFP/term) - Amp
- K081008 (LuxI) - Amp
- R0062 (pLux) - Amp
Nanodrop
Glycerol cultures
1125µl culture + 375µl glycerol (15%)
- J732006 (LacZ)
- B0031 (weak RBS)
- B0032 (medium RBS)
- B0034 (standard RBS)
- J31007 (TetA C)
- C0062 (LuxR w/o RBS)
- J37033 (RBS/LuxR) from the 2007 distribution plate
Aleksandra
Transformation of TOP10 cells with 2µl of the ligation product (ligation overnight 07/21-07/22) :
- pLux-RBS/GFP/term - Amp
- attc-term/attC/mRFP -Kan
- LuxI-term -Amp
→The attc-term/attC/mRFP and the LuxI-term plates have colonies! (07/23) Maybe our transformations worked.
Overnight cultures
2 falcons of 10 ml culture each for these parts :
- J732006 (LacZ)
- B0031 (weak RBS)
- B0032 (medium RBS)
- B0034 (standard RBS)
- J31007 (TetA C)
- C0062 (LuxR w/o RBS)
- J37033 (RBS/LuxR) from the 2007 distribution plate
- 23119 (Ps)
- J23110 (Pm)
- attC
- term/attC/mRFP
- B0015 (term)
- E0240 (RBS/GFP/term)
- K081008 (LuxI)
- R0062 (pLux)