Team:Warsaw/Calendar-Stage1/30 August 2010

From 2010.igem.org

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30.08.2010 Monday

Previous day: Set up liquid cultures of E.coli TOP10 with measured constructs [Ania P.]

We used a protocol described here

Testing the strenght of RBS [Ania P.,Cherry]

Preparation of samples by dilluting the bacteria cultures and examining them in flow cytometer (all according the protocol, but each RBS preformed from one independent culture)

Data analysis with Cell Quest and Microsoft Excel[Cherry]


legend:

30-08-2010.001- TOP10 without construct
30-08-2010.007- B0030
30-08-2010.008- B0031
30-08-2010.009- B0032
30-08-2010.010- B0033
30-08-2010.011- B0034
30-08-2010.012- J61100
30-08-2010.013- J61101
30-08-2010.014- J61107
30-08-2010.015- J61117
30-08-2010.016- J61127

cytometer plots:
FSC- Forward Scatter, cell size
SSC- Side Scatter, cell complexity
FL-1- FL-1 sensor, cell fluorescence



Cytometer data processed with Cell Quest:




Cell Quest data processed with Microsoft Excel:



geo mean- geometric mean of fluorescence of events in FL1
peak channel- nr of channel which recorded the biggest nr of events


partgeo meanpeak channel
allm1allm1
J6110022,1142,66154
J6110195,38362,431406
J6110761,84209,271228
J6111757,75171,531212
J6112721,3152,81152
B0030187,45913,8511074
B0031190,21907,6211154
B0032103,05406,11378
B00336,199,9111
B0034182,7999511333
partgeo mean %B0033peak channel %B0033
allm1allm1
J6110012,10%4,29%100,00%4,05%
J6110152,18%36,43%100,00%30,46%
J6110733,83%21,03%100,00%17,10%
J6111731,59%17,24%100,00%15,90%
J6112711,66%5,31%100,00%3,90%
B0030102,55%91,84%100,00%80,57%
B0031104,06%91,22%100,00%86,57%
B003256,38%40,81%100,00%28,36%
B00333,39%0,99%100,00%0,83%
B0034100,00%100,00%100,00%100,00%