Team:Warsaw/Calendar-Stage1/2 September 2010
From 2010.igem.org
02.09.2010 Wednesday
Previous day: Set up liquid cultures of E.coli TOP10 with measured constructs [Ania O.]
We used a protocol described here
Testing the strenght of RBS [Ania O.,Cherry]
Preparation of samples by dilluting the bacteria cultures and examining them in flow cytometer (all according the protocol
Data analysis with Cell Quest and Microsoft Excel[Cherry]
legend:
rbs.033- TOP10 without construct
rbs.004- 1) B0030
rbs.005- 2) B0030
rbs.006- 3) B0030
rbs.007- 1) B0031
rbs.008- 2) B0031
rbs.009- 3) B0031
rbs.010- 1) B0032
rbs.011- 2) B0032
rbs.012- 3) B0032
rbs.013- 1) B0033
rbs.014- 2) B0033
rbs.015- 3) B0033
rbs.003- 1) B0034
rbs.016- 2) B0034
rbs.017- 3) B0034
rbs.018- 1) J61100
rbs.019- 2) J61100
rbs.020- 3) J61100
rbs.021- 1) J61101
rbs.022- 2) J61101
rbs.023- 3) J61101
rbs.024- 1) J61107
rbs.025- 2) J61107
rbs.026- 3) J61107
rbs.027- 1) J61117
rbs.028- 2) J61117
rbs.029- 3) J61117
rbs.030- 1) J61127
rbs.031- 2) J61127
rbs.032- 3) 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
part | geo mean | peak channel | ||
all | m1 | all | m1 | |
top | 1,13 | 1,10 | 1,00 | 1,00 |
1) B0030 | 659,65 | 1134,44 | 1,00 | 1433,00 |
2) B0030 | 954,01 | 1596,95 | 1715,00 | 1715,00 |
3) B0030 | 633,97 | 1278,36 | 1,00 | 1485,00 |
mean | 749,21 | 1336,58 | 572,33 | 1544,33 |
1) B0031 | 181,89 | 292,73 | 1,00 | 327,00 |
2) B0031 | 364,01 | 623,60 | 1,00 | 723,00 |
3) B0031 | 174,81 | 293,39 | 1,00 | 283,00 |
mean | 240,24 | 403,24 | 1,00 | 444,33 |
1) B0032 | 106,08 | 164,61 | 1,00 | 245,00 |
2) B0032 | 28,73 | 65,32 | 1,00 | 67,00 |
3) B0032 | 145,17 | 220,27 | 1,00 | 237,00 |
mean | 93,33 | 150,07 | 1,00 | 183,00 |
1) B0033 | 4,93 | 5,69 | 1,00 | 7,00 |
2) B0033 | 3,09 | 2,83 | 1,00 | 1,00 |
3) B0033 | 3,44 | 3,82 | 1,00 | 4,00 |
mean | 3,82 | 4,11 | 1,00 | 4,00 |
1) B0034 | 357,81 | 1112,27 | 1,00 | 1197,00 |
2) B0034 | 705,27 | 1189,97 | 1,00 | 1433,00 |
3) B0034 | 605,00 | 1259,44 | 1,00 | 1654,00 |
mean | 556,03 | 1187,23 | 1,00 | 1428,00 |
1) J61100 | 13,29 | 23,49 | 1,00 | 35,00 |
2) J61100 | 11,32 | 19,18 | 1,00 | 22,00 |
3) J61100 | 20,40 | 29,91 | 1,00 | 45,00 |
mean | 15,00 | 24,19 | 1,00 | 34,00 |
1) J61101 | 114,13 | 212,72 | 1,00 | 283,00 |
2) J61101 | 117,92 | 221,76 | 1,00 | 264,00 |
3) J61101 | 223,47 | 370,24 | 1,00 | 437,00 |
mean | 151,84 | 268,24 | 1,00 | 328,00 |
1) J61107 | 59,50 | 121,76 | 1,00 | 143,00 |
2) J61107 | 82,37 | 161,98 | 1,00 | 237,00 |
3) J61107 | 62,99 | 132,14 | 1,00 | 205,00 |
mean | 68,29 | 138,63 | 1,00 | 195,00 |
1) J61117 | 13,54 | 23,18 | 1,00 | 36,00 |
2) J61117 | 24,57 | 40,39 | 1,00 | 52,00 |
3) J61117 | 14,27 | 24,28 | 1,00 | 35,00 |
mean | 17,46 | 29,28 | 1,00 | 41,00 |
1) J61127 | 45,87 | 124,24 | 1,00 | 212,00 |
2) J61127 | 46,82 | 120,56 | 1,00 | 143,00 |
3) J61127 | 37,56 | 98,11 | 1,00 | 93,00 |
mean | 43,42 | 114,30 | 1,00 | 149,33 |
part means | geo mean %B0034 | peak channel %B0034 | ||
all | m1 | all | m1 | |
top | 0% | 0% | 100% | 0% |
J61100 | 3% | 2% | 100% | 2% |
J61101 | 27% | 23% | 100% | 23% |
J61107 | 12% | 12% | 100% | 14% |
J61117 | 3% | 2% | 100% | 3% |
J61127 | 8% | 10% | 100% | 10% |
B0030 | 135% | 113% | 57233% | 108% |
B0031 | 43% | 34% | 100% | 31% |
B0032 | 17% | 13% | 100% | 13% |
B0033 | 1% | 0% | 100% | 0% |
B0034 | 100% | 100% | 100% | 100% |