Team:Stockholm/1 July 2010

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(Difference between revisions)
(New page: {{Stockholm/Top2}} = Andreas = == Preparation of competent BL21(DE3) cells == Competent cells were prepared as Team:Stockholm/1_July_2010)
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== Preparation of competent BL21(DE3) cells ==
== Preparation of competent BL21(DE3) cells ==
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Competent cells were prepared as [[Team:Stockholm/1_July_2010]]
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Competent cells were prepared as [https://2010.igem.org/Team:Stockholm/22_June_2010 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:
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==== Top10 ====
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*pSB1A3.BBa_J04450
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*pSB1C3.BBa_J04450
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*pSB1K3.BBa_J04450
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*pSB1AC3.BBa_J04450
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*pSB1AK3.BBa_J04450
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==== BL21(DE3) ====
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*pEX.SOD
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*pEX.yCCS
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*pMA.BBa_K157011
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== Isolation of requested parts ==
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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].

Revision as of 22:14, 1 July 2010


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].