Team:Stockholm/1 July 2010

From 2010.igem.org


Contents

Andreas

Preparation of competent BL21(DE3) cells

Competent cells were prepared as previously described.

Modifications
  • Growth temperature was changed to 37°C
  • Final OD(600) was 0.57

Preparation of LB agar plates with antibiotics

  • 50 ug/ml Km (20 plates)
  • 50 ug/ml Amp & 25 ug/ml Cm (10 plates)
  • 50 ug/ml Amp & 25 ug/ml Km (10 plates)

Plates were tested by plating with competent, non-transformed Top10 cells

Transformations

Several different transformations were performed. Top10 cells were transformed with pSB1x3 BioBrick vectors. BL21(DE3) cells were transformed with SOD- and yCCS-carrying pEX vectors as well as with the pMA vector to test competence:

Top10

  • pSB1A3.BBa_J04450
  • pSB1C3.BBa_J04450
  • pSB1K3.BBa_J04450
  • pSB1AC3.BBa_J04450
  • pSB1AK3.BBa_J04450

BL21(DE3)

  • pEX.SOD
  • pEX.yCCS
  • pMA.BBa_K157011

Isolation of requested parts

Our three requested parts BBa_J18930 (mCerulean; CFP), BBa_J18931 (mCitrine; YFP) and BBa_J18932 (mCherry; RFP) arrived as agar stabs. These were plated onto Amp plates according to [http://partsregistry.org/Help:Requesting_Parts_2009#Using_agar_stabs Registry instructions].