Difference between revisions of "Team:Aix-Marseille/InterLab"

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Description :
 
Description :
  
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We used plasmids containing different promoter from the 2017 distribution kit and transfect into E.coli. Before measuring fluorescence expression, we used LUDOX and fluorescein to calibrate the TECAN.
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We used plasmids containing different promoter from the 2017 distribution kit and transfected them into E.coli. Before measuring fluorescence expression, we used LUDOX and fluorescein to calibrate the TECAN.
−
We prepared a dilution series of fluorescein in 4 replicates and measure the fluorescence in   a 96 well plate in a plate reader. By measuring these in all standard modes,we generated a   standard curve of fluorescence for fluorescein concentration.We used this to correct cell based readings to an equivalent fluorescein concentration  and convert this into a concentration of GFP.
+
We prepared a dilution series of fluorescein in 4 duplicates and measure the fluorescence in a 96 well plate in a plate reader. By measuring these in all standard modes, we generated a standard curve of fluorescence for fluorescein concentration. We used this to correct cell based readings to an equivalent fluorescein concentration  and convert this into a concentration of GFP.
  
  

Revision as of 14:51, 29 September 2017

InterLab


T--Aix-Marseille--fluo interlab.jpeg

Description :

We used plasmids containing different promoter from the 2017 distribution kit and transfected them into E.coli. Before measuring fluorescence expression, we used LUDOX and fluorescein to calibrate the TECAN. We prepared a dilution series of fluorescein in 4 duplicates and measure the fluorescence in a 96 well plate in a plate reader. By measuring these in all standard modes, we generated a standard curve of fluorescence for fluorescein concentration. We used this to correct cell based readings to an equivalent fluorescein concentration and convert this into a concentration of GFP.


Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 156 165 167 177 157 159 153 153 147
Colony 1, Replicate 2 155 161 166 161 151 160 156 158 154
Colony 1, Replicate 3 158 157 164 152 144 159 151 155 147
Colony 1, Replicate 4 157 161 165 155 150 156 155 155 155
Colony 2, Replicate 1 158 159 163 178 154 158 158 153 152
Colony 2, Replicate 2 89 104 97 112 93 99 100 90 91
Colony 2, Replicate 3 94 103 99 109 93 103 99 97 89
Colony 2, Replicate 4 92 102 101 80 89 95 94 97 90


Hour 2: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 102 132 124 121 98 117 112 99 93
Colony 1, Replicate 2 101 126 120 125 99 109 118 96 93
Colony 1, Replicate 3 98 126 121 116 98 113 115 91 84
Colony 1, Replicate 4 99 128 122 118 91 111 115 93 92
Colony 2, Replicate 1 105 133 118 139 101 111 116 102 93
Colony 2, Replicate 2 99 128 113 139 103 110 114 104 95
Colony 2, Replicate 3 100 130 110 131 102 109 113 102 97
Colony 2, Replicate 4 96 132 116 133 101 116 112 99 97


Hour 4: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 109 228 168 250 105 147 152 105 93
Colony 1, Replicate 2 112 236 173 254 101 146 166 106 92
Colony 1, Replicate 3 112 243 174 259 101 154 162 108 85
Colony 1, Replicate 4 114 242 171 255 101 153 149 108 92
Colony 2, Replicate 1 103 224 157 262 99 151 156 106 102
Colony 2, Replicate 2 109 239 165 274 104 157 161 108 93
Colony 2, Replicate 3 110 239 161 263 105 162 159 109 91
Colony 2, Replicate 4 108 241 167 272 106 166 169 112 95


Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 148 362 348 481 147 343 211 142 120
Colony 1, Replicate 2 156 378 351 510 137 347 219 148 120
Colony 1, Replicate 3 150 382 318 497 138 355 218 153 120
Colony 1, Replicate 4 154 385 354 479 138 358 210 151 130
Colony 2, Replicate 1 149 354 336 493 136 375 210 150 130
Colony 2, Replicate 2 149 391 365 564 146 399 193 147 128
Colony 2, Replicate 3 154 367 355 507 140 416 210 149 130
Colony 2, Replicate 4 153 385 359 560 148 417 232 151 138