IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/07/21

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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:
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 big colony plate 1
 +
-one small colony plate 1
-one small colony plate 1
 +
-one big colony plate 2
-one big colony plate 2
 +
-one small colony plate 2
-one small colony plate 2

Revision as of 14:10, 22 July 2010

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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)


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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