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Description : | Description : | ||
− | We used plasmids containing different promoter from the 2017 distribution kit and | + | 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 | + | 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
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 |