Tecan Parameters
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SAFIR E II; Serialnumber:807003897; Firm w are:V 2.10 12/2007 Safire2; XFLUO R 4SAFIR EIIVersion:V 4.62n Date: 28/7/09 Time: 20:23 Measurem entm ode: Fluorescence Excitation w avelength: 488 nm Em ission w avelength: 575 nm Excitation bandw idth: 20 nm Em ission bandw idth: 20 nm G ain (M anual): 50 N um berofreads: 20 FlashM ode: H igh sensitivity Integration tim e: 40 µs Lag tim e: 0 µs Plate definition file: CO S96ft.pdf Z-Position (M anual): 5100 µm TargetTemperature: 37 °C C urrentTem perature: 37 °C R awdata (R FU ) Temperature: 37 °C Tecan Parameters
description
Tecan Parameters. OD normalized Data. cel5B. Orange = induced parts green = uninduced parts light pink = induced pos EC100D pink = uninduced pos EC100D light green = induced neg EC100D blue = uninduced neg EC100D gray = induced pos DH10B. cel5B. After 3 washes. - PowerPoint PPT Presentation
Transcript of Tecan Parameters
SAFIRE II; Serial number: 807003897; Firmware: V 2.10 12/2007 Safire2; XFLUOR4SAFIREII Version: V 4.62nDate: 28/7/09Time: 20:23
Measurement mode: FluorescenceExcitation wavelength: 488 nmEmission wavelength: 575 nmExcitation bandwidth: 20 nmEmission bandwidth: 20 nmGain (Manual): 50Number of reads: 20FlashMode: High sensitivityIntegration time: 40 µsLag time: 0 µsPlate definition file: COS96ft.pdfZ-Position (Manual): 5100 µmTarget Temperature: 37 °CCurrent Temperature: 37 °C
Rawdata (RFU) Temperature: 37 °C
Tecan Parameters
OD normalized Data
<> 1 2 3 4 5 6 7 8 9 10 11 12A 314.5236 328.4672 279.1878 279.7203 271.4165 293.8634 305.2065 275.9197 25885.34 120 136.6224 131.2741B 330.4179 321.7158 293.3563 288.6406 317.757 281.6901 279.5311 288.1356 25958.11 141.0203 122.4036 119.0934C 314.9606 270.027 241.4045 256.8093 250.8091 252.9762 234.0094 240 28603.19 156.3218 128.866 142.8571D 316.6227 270.2703 257.6891 279.6053 288.9667 305.1439 272.4359 288.2096 3295.113 160.8363 132.1586 155.2644E 281.2242 220.2073 288.3031 241.8745 240.1922 322.8621 260.5863 284.3217 3544.382 124.5621 125.138 151.2287F 280.0659 258.6904 308.37 247.9942 254.7771 288.2883 330.3571 316.7421 615.3846 112.4385 143.4978 161.6063G 551.0204 684.2105 405.6162 492.0914 552.381 424.2424 418.5351 299.7602 387.2752 319.9052 370.8282 244.7869H 708.6614 748.6631 678.8512 786.5169 675.3247 704.607 750.6702 737.7049 794.5205 725.8065 680.6283 795.756
cel5B
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Orange = induced parts green = uninduced parts light pink = induced pos EC100D pink = uninduced pos EC100D light green = induced neg EC100D blue = uninduced neg EC100D gray = induced pos DH10B
cel5B
After 3 washes
<> 1 2 3 4 5 6 7 8 9 10 11 12A 349.7758 345.1408 324.9097 349.7758 25605 206.2919 132.9243 121.4446 765.1715 137.9022 129.9213 150.9099B 335.4488 357.7982 333.0487 367.3095 23403.6 205.5165 132.1253 148.6989 714.2857 147.2684 144.1678 133.9884C 327.4559 304.6595 299.1773 298.7198 26244.3 206.0222 157.2052 181.2268 430.622 166.4289 157.7438 153.1059D 808.9888 534.9794 545.4545 656.0636 589.5197 507.0994 599.0783 486.3813 500 585.6833 625 698.324E 339.1304 557.8512 347.9853 293.884 315.7895 287.4903 239.8401 319.1489 306.2117 304.3478 189.8409 329.6703F 335.0084 654.4503 316.6667 294.599 291.8587 304.0263 237.6103 315.4876 275.3623 305.7851 176.8489 290.5983G 312 603.0151 270.2703 305.7851 318.4713 286.6242 266.3623 272.1088 283.5696 295.9029 191.3876 261.8542H 612.2449 687.9607 716.1804 740.7407 510.5634 634.9206 743.5897 788.0435 746.6667 789.4737 700.4831 731.7073
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cel9A – OD normalized
Orange = induced parts green = uninduced parts light pink = induced pos EC100D pink = uninduced pos EC100D light green = induced neg EC100D blue = uninduced neg EC100D gray = induced pos DH10B
cel9A after 3 washes
Orange = induced parts green = uninduced parts light pink = induced pos EC100D pink = uninduced pos EC100D light green = induced neg EC100D blue = uninduced neg EC100D gray = induced pos DH10B
gliadin – OD normalized<> 1 2 3 4 5 6 7 8 9 10 11 12A 319.2338 289.5323 154.265 257.766 21218.57 188.9339 167.1309 190.9641 4093.445 157.8255 150.7321 153.3071B 289.103 239.3038 148.3813 260.0958 19814.6 173.8375 161.9621 184.5018 4129.727 157.6873 151.7122 167.5485C 287.226 252.1614 172.978 239.1171 296.3776 182.6045 170.3163 200.4773 388.0983 182.6259 167.5443 170.052D 280.0295 291.9708 271.3816 308.7886 300.2309 167.5702 266.5706 294.8022 260.223 253.365 268.7802 304.3868E 260.989 300.8596 309.5975 289.6341 291.411 161.3647 257.6271 266.5706 259.8315 284.5528 270.8333 302.0134F 283.6879 283.0189 295.7198 333.8633 319.3146 168.6183 299.3197 286.5995 295.9502 324.2321 280.236 334.2367G 560 668.2578 428.1346 498.2206 534.07 400 398.8604 314.9606 358.5147 305.9072 328.4672 250.8961H 676.6917 687.2038 639.3862 734.1772 681.2652 710.6599 731.0705 771.2766 784.3137 798.8981 721.393 815.7895
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Gliadin scFv after 3 washes
Needle – OD normalized
Orange = induced parts green = uninduced parts light pink = induced pos EC100D pink = uninduced pos EC100D light green = induced neg EC100D blue = uninduced neg EC100D gray = induced pos DH10B
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<> 1 2 3 4 5 6 7 8 9 10 11 12A 238.6635 252.0516 229.682 232.5581 177.8656 223.0259 226.8761 238.0952 21741.63 702.3411 170.4303 203.1176B 196.8912 232.1429 225.2519 198.3664 230.2821 204.0816 217.1492 221.0992 20004.04 721.393 205.2335 233.1887C 261.8454 242.464 222.4758 211.2285 212.8852 203.8505 200.4334 198.7852 22187.3 412.3711 227.9202 214.8847D 234.7919 157.7287 404.4118 446.4286 486.3813 468.75 539.3258 146.7748 187.5977 210.0162 158.6074 201.4099E 228.0311 203.6442 400 479.1667 500.9634 500 460.6526 210.5832 176.416 206.1311 207.6843 193.3702F 256.0976 204.4199 452.7559 658.2278 605.3269 516.8539 585.4801 205.7143 202.4507 205.7416 204.1943 218.1818G 568.6275 640.7323 465.4655 514.1844 543.4783 412.6547 403.8997 357.5548 358.5147 299.1027 339.9766 275.8007H 759.4937 732.3232 723.5142 806.4516 779.2208 721.6495 719.7943 818.7135 712.4011 769.2308 777.2021 753.2468
Needle scFv after 3 washes
Some Observations
• pBdr052 has toxic part under pBad
contructs in EC100D and pBdr052positive control in EC100D and pBca9145negative control in EC100D and pBca9495positive control in DH10B and pBca9145
neg induced induced constructs166.4698 298.521 260.3325 158.5414 252.3263 274.9021 291.9498 292.233 310.762 303.6522 165.8511 274.5058 282.6574 272.0016 287.3833 273.2832 313.5456
induced constructs/neg induced1.793244 1.563842 0.952374 1.515748 1.651363 1.75377 1.755471 1.866777 1.824068 0.996283 1.648983 1.69795 1.633939 1.726339 1.641638 1.883498
induced constructs298.521 260.3325 158.5414
uninduced constructs166.0462 156.6629 163.6358induced constructs/uninduced constructs1.797819 1.661737 0.968867
