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Team:Stockholm/1 July 2010
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(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 == | ||
| - | Competent cells were prepared as [ | + | 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: | ||
| + | ==== 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]. | ||
Revision as of 22:14, 1 July 2010
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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].
