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 and grown on agar plates with appropriate antibiotic(s). BL21(DE3) cells were transformed with SOD- and yCCS-carrying pEX vectors as well as with the pMA vector to test cell competence and grown on Amp plates:

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].

Maya

1 Microarrays - reliability - How reliable is the protein regulation profile in the article "Transcriptional profiling of melanocytes..."

2 How should one priorate different types of protein-protein interactions? Focus on experimental (not predictive ) data and focus first of all at physical PPI second at co expression and co localisation. Skip the rest.

To improve PPI reliability

Find a good coverage from many primary databases. -How many and with?
What experiments were used? If the same result is dereved from many different experiments then the PPI is more reliable.
Look at 3D structures...?

Compare with canonical pathways to get a better insight in the functional roles of the proteins in the network.

Team:Stockholm/1 July 2010

From 2010.igem.org

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