* Report Run Properties Name Praktikum 20180423 File C:\Users\Milan\aTubeCatcher\Documents\Bio Molecular Systems\micPCR\Assays\Praktikum 20180423.micrun Signature Valid Status Completed Operator Milan Started 23.4.2018 14:39:55 Completed 23.4.2018 16:04:21 Event Log Time Priority User Message 23.4.2018 14:39:55 Information Milan Run started via Serial on "mic M0000163" S/N M0000163 F/Wv2.24 S/Wv2.6.4 23.4.2018 14:54:35 Information Milan Autogain completed for Green using Sample: 2, Detector gain: 3x, LED power: 364, Scale: 1,02 23.4.2018 14:54:37 Information Milan Autogain completed for Yellow using Sample: 2, Detector gain: 2x, LED power: 392, Scale: 0,97 23.4.2018 16:04:21 Information Milan Run complete 23.4.2018 16:07:33 Information Milan Cycling "VZV" added Method changed from "LinRegPCR" to "Dynamic" Samples selected: 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22 23.4.2018 16:07:33 Information Milan Cycling "IC" added Method changed from "LinRegPCR" to "Dynamic" 23.4.2018 16:07:33 Information Milan AbsoluteQuantification "VZV" added Cycling "VZV" added Method changed from "LinRegPCR" to "Dynamic" Generated by micPCR version 2.6.4 at 23.4.2018 16:07:37 Page 1of12 Report Samples Praktikum 20180423 Well Name Type Std. Cone. (Copies/|jL) Groups Assay 1 Kalibrátor 10e1 Standard 10 Varicellazoster Virus_praktikum 2 Kalibrátor 10e2 Standard 100 Varicellazoster Virus_praktikum 3 Kalibrátor 10e3 Standard 1000 Varicellazoster Virus_praktikum 4 Kalibrátor 10e4 Standard 10000 Varicellazoster Virus_praktikum 5 Kalibrátor 10e5 Standard 100000 Varicellazoster Virus_praktikum 6 NC Negative Control Varicellazoster Virus_praktikum 7 451182 Unknown Varicellazoster Virus_praktikum 8 451182 Unknown Varicellazoster Virus_praktikum 9 RK Unknown Varicellazoster Virus_praktikum 10 RK Unknown Varicellazoster Virus_praktikum 11 MČ Unknown Varicellazoster Virus_praktikum 12 MČ Unknown Varicellazoster Virus_praktikum 13 LIV Unknown Varicellazoster Virus_praktikum 14 LIV Unknown Varicellazoster Virus_praktikum 15 JS Unknown Varicellazoster Virus_praktikum 16 JS Unknown Varicellazoster Virus_praktikum 17 JH Unknown Varicellazoster Virus_praktikum 18 JH Unknown Varicellazoster Virus_praktikum 19 KK Unknown Varicellazoster Virus_praktikum 20 KK Unknown Varicellazoster Virus_praktikum 21 NH Unknown Varicellazoster Virus_praktikum 22 NH Unknown Varicellazoster Virus_praktikum 23 Unknown Varicellazoster Virus_praktikum 24 Unknown 25 Unknown 26 Unknown 27 Unknown 28 Unknown 29 Unknown Generated by micPCR version 2.6.4 at 23.4.2018 16:07:38 Page 2 of 12 Report Praktikum 20180423 Well Name Type Std. Cone. (Copies/|jL) Groups Assay 30 Unknown 31 Unknown 32 Unknown 33 Unknown 34 Unknown 35 Unknown 36 Unknown 37 Unknown 38 Unknown 39 Unknown 40 Unknown 41 Unknown 42 Unknown 43 Unknown 44 Unknown 45 Unknown 46 Unknown 47 Unknown 48 Unknown Generated by micPCR version 2.6.4 at 23.4.2018 16:07:38 Page 3 of 12 Report Run Profile Praktikum 20180423 Hold Steps Activation at 37°C for 2:00 Hold at 95°C for 10:00 Cycling 1) 95°Cfor5s 40 cycles 2) 60°C for 40s acquiring on Green, Yellow 3) 72°Cfor20s Generated by micPCR version 2.6.4 at 23.4.2018 16:07:40 Page 4 of 12 Report Cycling: VZV Praktikum 20180423 Target VaricellaZoster Virus_praktikum —► VZV Normalisation Dynamic Exclusion Extensive with fluorescence cutoff of 5% Threshold 0,155 (Automatic) starting at cycle 1 ■J d ■B ■J O -3 0.1 E i_ o 0.01 0.001 Well Colour Sample Name Cq Efficiency Efficiency R2 Result 1 ■ Kalibrátor 10e1 34,11 0,70 0,99749 2 ■ Kalibrátor 10e2 32,40 0,89 0,99911 3 Kalibrátor 10e3 28,42 1,07 0,99922 4 Kalibrátor 10e4 24,80 1,17 0,99995 5 Kalibrátor 10e5 21,95 1,09 0,99981 6 NC - - - Excluded 7 451182 33,53 0,59 0,99685 8 451182 34,84 0,57 0,97314 9 RK - - - Excluded 10 RK - - - Excluded 11 ■ MČ - - - Excluded 12 ■ MČ - - - Excluded 13 ■ LIV 30,51 0,72 0,99887 14 ■ LIV 31,03 0,90 0,99923 15 ■ JS 32,29 0,71 0,99875 16 ■ JS 31,82 0,83 0,99723 Generated by micPCR version 2.6.4 at 23.4.2018 16:07:42 Page 5 of 12 Report Praktikum 20180423 Well Colour Sample Name Cq Efficiency Efficiency R2 Result 17 ■ JH 32,41 0,82 0,99596 18 ■ JH 31,94 0,73 0,99729 19 ■ KK 36,66 0,51 0,99299 20 ■ KK 34,86 0,62 0,98977 21 NH 35,88 0,46 0,99774 22 NH 34,49 0,71 0,99105 Generated by micPCR version 2.6.4 at 23.4.2018 16:07:42 Page 6 of 12 Report Cycling: IC Praktikum 20180423 Target VaricellaZoster Virus_praktikum —► IC Normalisation Dynamic Exclusion Extensive with fluorescence cutoff of 5% Threshold 0,733 (Automatic) starting at cycle 1 i: 0.001 ■ ■ /i \, 1 ill 5 10 15 20 25 35 40 Cycle Well Colour Sample Name Cq Efficiency Efficiency R2 Result 1 ■ Kalibrátor 10e1 29,02 0,99 0,99854 2 ■ Kalibrátor 10e2 28,77 0,94 0,99868 3 Kalibrátor 10e3 28,98 1,03 0,99787 4 Kalibrátor 10e4 28,61 0,93 0,99647 5 Kalibrátor 10e5 28,51 0,67 0,99531 6 NC 29,30 0,97 0,99906 7 451182 29,00 0,93 0,99914 8 451182 28,77 1,00 0,99796 9 RK 29,05 0,95 0,99847 10 RK 29,06 0,96 0,99856 11 ■ MČ 29,25 0,96 0,99856 12 ■ MČ 29,21 0,98 0,99837 13 ■ LIV 29,01 0,95 0,99849 14 ■ LIV 29,00 0,97 0,99778 15 ■ JS 29,04 0,94 0,99806 16 ■ JS 29,02 0,96 0,99727 Generated by micPCR version 2.6.4 at 23.4.2018 16:07:42 Page 7 of 12 Report Praktikum 20180423 Well Colour Sample Name Cq Efficiency Efficiency R2 Result 17 ■ JH 29,05 0,96 0,99762 18 ■ JH 29,22 0,98 0,99859 19 ■ KK 29,07 0,97 0,99796 20 ■ KK 29,03 0,97 0,99815 21 NH 29,27 0,96 0,99871 22 NH 29,07 0,96 0,99813 23 29,29 1,00 0,99877 Generated by micPCR version 2.6.4 at 23.4.2018 16:07:43 Page 8 of 12 Report Absolute Quantification: VZV Praktikum 20180423 Standard Curve Equation y = -3,19 x+ 37,91 Efficiency 1,06 R2 0,9879 Standards Results Well Sample Name Cq Given Concentration (Copies/uL) Calculated Concentration (Copies/uL) Variation 1 Kalibrátor 10e1 34,11 10 15,51 55,11% 2 Kalibrátor 10e2 32,40 100 53,38 46,62% 3 Kalibrátor 10e3 28,42 1000 942,8 5,72% 4 Kalibrátor 10e4 24,80 1E+04 1.278E+04 27,82% 5 Kalibrátor 10e5 21,95 1E+05 1.002E+05 0,23% Sample Results Well Sample Name Cq Calculated Concentration (Copies/uL) 6 NC - - 7 451182 33,53 23,52 8 451182 34,84 9,129 9 RK - - 10 RK - - Generated by micPCR version 2.6.4 at 23.4.2018 16:07:43 Page 9 of 12 Report Praktikum 20180423 Well Sample Name Cq Calculated Concentration (Copies/uL) 11 MC - - 12 MC - - 13 LIV 30,51 208,4 14 LIV 31,03 142,9 15 JS 32,29 57,68 16 JS 31,82 81 17 JH 32,41 52,81 18 JH 31,94 74,04 19 KK 36,66 2,463 20 KK 34,86 9,014 21 NH 35,88 4,312 22 NH 34,49 11,74 23 - - Generated by micPCR version 2.6.4 at 23.4.2018 16:07:43 Page 10 of 12 Report Praktikum 20180423 Absolute Quantification: VZV (Cycling) Target VaricellaZoster Virus_praktikum —► VZV Normalisation Dynamic Exclusion Extensive with fluorescence cutoff of 5% Threshold 0,155 (Automatic) starting at cycle 1 ■J d ■B ■J O -3 0.1 E i_ o 0.01 0.001 Well Colour Sample Name Cq Efficiency Efficiency R2 Result 1 ■ Kalibrátor 10e1 34,11 0,70 0,99749 2 ■ Kalibrátor 10e2 32,40 0,89 0,99911 3 Kalibrátor 10e3 28,42 1,07 0,99922 4 Kalibrátor 10e4 24,80 1,17 0,99995 5 Kalibrátor 10e5 21,95 1,09 0,99981 6 NC - - - Excluded 7 451182 33,53 0,59 0,99685 8 451182 34,84 0,57 0,97314 9 RK - - - Excluded 10 RK - - - Excluded 11 ■ MČ - - - Excluded 12 ■ MČ - - - Excluded 13 ■ LIV 30,51 0,72 0,99887 14 ■ LIV 31,03 0,90 0,99923 15 ■ JS 32,29 0,71 0,99875 16 ■ JS 31,82 0,83 0,99723 Generated by micPCR version 2.6.4 at 23.4.2018 16:07:43 Page 11 of 12 Report Praktikum 20180423 Well Colour Sample Name Cq Efficiency Efficiency R2 Result 17 ■ JH 32,41 0,82 0,99596 18 ■ JH 31,94 0,73 0,99729 19 ■ KK 36,66 0,51 0,99299 20 ■ KK 34,86 0,62 0,98977 21 NH 35,88 0,46 0,99774 22 NH 34,49 0,71 0,99105 23 - - - Excluded Generated by micPCR version 2.6.4 at 23.4.2018 16:07:43 Page 12 of 12