Luận văn : BƯỚC ĐẦU XÂY DỰNG QUY TRÌNH ĐỊNH LƯỢNG CÁC SẢN PHẨM BIẾN ĐỔI GEN BẰNG PHƯƠNG PHÁP REAL-TIME PCR part 10 ppsx

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104 104 Gen tham chiếu: PCR Standard Curve Report with PCR Amp Cycle Graph PCR Base Line Subtracted Data Current Date: 19-Aug-05 12:05 PM Data generated on: 22-Jul-05 at 06:15 PM. Optical data file name: Data 22-Jul-05 1815-Truong-35S-dot 1-sua2.odm Plate Setup file used: GMO-35S.psm Protocol file used: GMO-35S.tmo Sample volume: 25.00 ul Hot Start? No Well factor collection: Experimental Plate Comments Protocol Cycle 1: ( 1X) Step 1: 95.0ºC for 10:00 Cycle 2: ( 45X) Step 1: 95.0ºC for 00:15 Step 2: 60.0ºC for 01:00 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC HOLD PCR Amp/Cycle Graph for FAM-490 Standard Curve Graph for FAM-490 105 105 Data Analysis Parameters Calculated threshold has been replaced by the user selected threshold 67.5. User selected baseline cycles are 2 to 20. Data analysis window is set at 95.00% of a cycle, centered at end of the cycle. Weighted Mean digital filtering has been applied. Global filtering is off. Standard Curve Spreadsheet Data for FAM-490 Units: copy number Type Identifier Rep Ct Log SQ SQ SQ Ct Ct Set SQ Mean SD Mean SD Point B02 Standard Stan Corn-Ref 1 28.29 4.612 4.10E+04 4.10E+04 N/A 28.29 N/A B04 Standard Stan Corn-Ref 2 30.26 3.709 5.12E+03 5.12E+03 N/A 30.26 N/A B06 Standard Stan Corn-Ref 3 32.97 2.806 6.40E+02 6.40E+02 N/A 32.97 N/A B08 Standard Stan Corn-Ref 4 35.43 1.903 8.00E+01 8.00E+01 N/A 35.43 N/A B11 Unknown Bt 9 28.93 4.308 2.03E+04 2.03E+04 N/A 28.93 N/A Đợt 3: Định lƣợng bắp chuyển gen Bt 176 1% (lần 2) Promoter 35S: PCR Standard Curve Report with PCR Amp Cycle Graph PCR Base Line Subtracted Curve Fit Data Current Date: 19-Aug-05 12:17 PM Data generated on: 16-Aug-05 at 04:58 PM. Optical data file name: Data 16-Aug-05 1658 taqman dot 3-35S -Bt176.odm Plate Setup file used: 35S TAQMAN DOT 3.psm Protocol file used: GMO-35S.tmo Sample volume: 25.00 ul Hot Start? No Well factor collection: Experimental Plate Comments Protocol Cycle 1: ( 1X) Step 1: 95.0ºC for 10:00 Cycle 2: ( 45X) Step 1: 95.0ºC for 00:15 Step 2: 60.0ºC for 01:00 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC HOLD PCR Amp/Cycle Graph for FAM-490 106 106 Standard Curve Graph for FAM-490 Data Analysis Parameters Calculated threshold has been replaced by the user selected threshold 21.8. Per-well baseline cycles have been determined automatically. Data analysis window is set at 95.00% of a cycle, centered at end of the cycle. Weighted Mean digital filtering has been applied. Global filtering is off. Standard Curve Spreadsheet Data for FAM-490 Units: copy number Type Identifier Rep Ct Log SQ SQ SQ Ct Ct Set SQ Mean SD Mean SD Point E02 Standard ST 35S 5 23.85 3.709 5.12E+03 5.12E+03 N/A 23.85 N/A E03 Standard ST 35S 6 26.55 2.806 6.40E+02 6.40E+02 N/A 26.55 N/A E04 Standard ST 35S 7 29.13 1.903 8.00E+01 8.00E+01 N/A 29.13 N/A E05 Standard ST 35S 8 31.67 1.000 1.00E+01 1.00E+01 N/A 31.67 N/A F02 Unknown Bt 19 28.56 2.092 1.24E+02 1.24E+02 N/A 28.56 N/A Gen tham chiếu: PCR Standard Curve Report with PCR Amp Cycle Graph PCR Base Line Subtracted Data Current Date: 19-Aug-05 12:22 PM Data generated on: 16-Aug-05 at 04:58 PM. Optical data file name: Data 16-Aug-05 1658 taqman dot 3-DC Bt 176.odm Plate Setup file used: 35S TAQMAN DOT 3.psm Protocol file used: GMO-35S.tmo Sample volume: 25.00 ul 107 107 Hot Start? No Well factor collection: Experimental Plate Comments Protocol Cycle 1: ( 1X) Step 1: 95.0ºC for 10:00 Cycle 2: ( 45X) Step 1: 95.0ºC for 00:15 Step 2: 60.0ºC for 01:00 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC HOLD PCR Amp/Cycle Graph for FAM-490 Standard Curve Graph for FAM-490 Data Analysis Parameters Calculated threshold has been replaced by the user selected threshold 21.2. User selected baseline cycles are 2 to 20. Data analysis window is set at 95.00% of a cycle, centered at end of the cycle. Weighted Mean digital filtering has been applied. Global filtering is off. Standard Curve Spreadsheet Data for FAM-490 Units: copy number Type Identifier Rep Ct Log SQ SQ SQ Ct Ct Set SQ Mean SD Mean SD Point B02 Standard ST CR 1 23.31 4.612 4.10E+04 4.10E+04 N/A 23.31 N/A B03 Standard ST CR 2 25.71 3.709 5.12E+03 5.12E+03 N/A 25.71 N/A B04 Standard ST CR 3 27.04 2.806 6.40E+02 6.40E+02 N/A 27.04 N/A 108 108 B05 Standard ST CR 4 30.64 1.903 8.00E+01 8.00E+01 N/A 30.64 N/A C02 Unknown Bt 3 24.51 4.097 1.25E+04 1.25E+04 N/A 24.51 N/A Đợt 4: Độ nhạy của phản ứng PCR Standard Curve Report with PCR Amp Cycle Graph PCR Base Line Subtracted Curve Fit Data Current Date: 30-Aug-05 06:46 AM Data generated on: 22-Jul-05 at 06:15 PM. Optical data file name: Data 22-Jul-05 1815-Truong-35S-dot 1-do nhay.odm Plate Setup file used: GMO-35S.psm Protocol file used: GMO-35S.tmo Sample volume: 25.00 ul Hot Start? No Well factor collection: Experimental Plate Comments Protocol Cycle 1: ( 1X) Step 1: 95.0ºC for 10:00 Cycle 2: ( 45X) Step 1: 95.0ºC for 00:15 Step 2: 60.0ºC for 01:00 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC HOLD PCR Amp/Cycle Graph for FAM-490 Standard Curve Graph for FAM-490 109 109 Data Analysis Parameters Calculated threshold has been replaced by the user selected threshold 13.7. User selected baseline cycles are 2 to 24. Data analysis window is set at 95.00% of a cycle, centered at end of the cycle. Weighted Mean digital filtering has been applied. Global filtering is off. Standard Curve Spreadsheet Data for FAM-490 Units: copy number Type Identifier Rep Ct Log SQ SQ SQ Ct Ct Set SQ Mean SD Mean SD Point E02 Standard Stan Corn-Ref 5 25.73 3.709 5.12E+03 5.12E+03 N/A 25.73 N/A E04 Standard Stan Corn-Ref 6 28.57 2.806 6.40E+02 6.40E+02 N/A 28.57 N/A E06 Standard Stan Corn-Ref 7 31.65 1.903 8.00E+01 8.00E+01 N/A 31.65 N/A E08 Standard Stan Corn-Ref 8 33.12 1.000 1.00E+01 1.00E+01 N/A 33.12 N/A F08 Unknown Mau 1 copy 4 36.22 0.047 1.11E+00 1.25E+00 1.91E-01 36.09 1.87E-01 F10 Unknown Mau 1 copy 4 35.96 0.141 1.38E+00 1.25E+00 1.91E-01 36.09 1.87E-01 Đợt 5: Định lƣợng các mẫu thí nghiệm (lần 1) Promoter 35S: PCR Standard Curve Report with PCR Amp Cycle Graph PCR Base Line Subtracted Curve Fit Data Current Date: 11-Aug-05 03:43 PM Data generated on: 22-Jul-05 at 06:15 PM. Optical data file name: Data 22-Jul-05 1815-Truong-35S-dot 1-sua.odm Plate Setup file used: GMO-35S.psm Protocol file used: GMO-35S.tmo Sample volume: 25.00 ul Hot Start? No Well factor collection: Experimental Plate Comments Protocol 110 110 Cycle 1: ( 1X) Step 1: 95.0ºC for 10:00 Cycle 2: ( 45X) Step 1: 95.0ºC for 00:15 Step 2: 60.0ºC for 01:00 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC HOLD PCR Amp/Cycle Graph for FAM-490 Standard Curve Graph for FAM-490 Data Analysis Parameters Calculated threshold has been replaced by the user selected threshold 52.2. User selected baseline cycles are 2 to 24. Data analysis window is set at 95.00% of a cycle, centered at end of the cycle. Weighted Mean digital filtering has been applied. Global filtering is off. Standard Curve Spreadsheet Data for FAM-490 Units: copy number Type Identifier Rep Ct Log SQ SQ SQ Ct Ct Set SQ Mean SD Mean SD Point E02 Standard Stan Corn-Ref 5 27.43 3.709 5.12E+03 5.12E+03 N/A 27.43 N/A E04 Standard Stan Corn-Ref 6 30.76 2.806 6.40E+02 6.40E+02 N/A 30.76 N/A E06 Standard Stan Corn-Ref 7 33.65 1.903 8.00E+01 8.00E+01 N/A 33.65 N/A E08 Standard Stan Corn-Ref 8 35.89 1.000 1.00E+01 1.00E+01 N/A 35.89 N/A F02 Unknown B1 5 27.38 3.808 6.42E+03 5.94E+03 6.84E+02 27.49 1.57E-01 F04 Unknown B1 5 27.61 3.737 5.46E+03 5.94E+03 6.84E+02 27.49 1.57E-01 F06 Unknown B2 6 N/A N/A N/A N/A N/A N/A N/A F08 Unknown B2 6 29.96 2.985 9.65E+02 9.65E+02 N/A 29.96 N/A G02 Unknown B7 7 27.71 3.704 5.06E+03 4.90E+03 2.23E+02 27.75 6.18E-02 G04 Unknown B7 7 27.80 3.676 4.75E+03 4.90E+03 2.23E+02 27.75 6.18E-02 111 111 Gen tham chiếu: PCR Standard Curve Report with PCR Amp Cycle Graph PCR Base Line Subtracted Curve Fit Data Current Date: 11-Aug-05 04:02 PM Data generated on: 22-Jul-05 at 06:15 PM. Optical data file name: Data 22-Jul-05 1815-Truong-35S-dot 1-2.odm Plate Setup file used: GMO-35S.psm Protocol file used: GMO-35S.tmo Sample volume: 25.00 ul Hot Start? No Well factor collection: Experimental Plate Comments Protocol Cycle 1: ( 1X) Step 1: 95.0ºC for 10:00 Cycle 2: ( 45X) Step 1: 95.0ºC for 00:15 Step 2: 60.0ºC for 01:00 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC HOLD PCR Amp/Cycle Graph for FAM-490 Standard Curve Graph for FAM-490 112 112 Data Analysis Parameters Calculated threshold has been replaced by the user selected threshold 72.9. User selected baseline cycles are 2 to 20. Data analysis window is set at 95.00% of a cycle, centered at end of the cycle. Weighted Mean digital filtering has been applied. Global filtering is off. Standard Curve Spreadsheet Data for FAM-490 Units: copy number Type Identifier Rep Ct Log SQ SQ SQ Ct Ct Set SQ Mean SD Mean SD Point B02 Standard Stan Corn-Ref 1 28.55 4.612 4.10E+04 4.10E+04 N/A 28.55 N/A B04 Standard Stan Corn-Ref 2 30.62 3.709 5.12E+03 5.12E+03 N/A 30.62 N/A B06 Standard Stan Corn-Ref 3 33.22 2.806 6.40E+02 6.40E+02 N/A 33.22 N/A B08 Standard Stan Corn-Ref 4 35.84 1.903 8.00E+01 8.00E+01 N/A 35.84 N/A C02 Unknown B1 1 27.69 4.870 7.42E+04 9.61E+04 3.09E+04 27.42 3.86E-01 C04 Unknown B1 1 27.14 5.072 1.18E+05 9.61E+04 3.09E+04 27.42 3.86E-01 C06 Unknown B2 2 37.03 1.420 2.63E+01 8.38E+01 8.13E+01 36.04 1.40E+00 C08 Unknown B2 2 35.06 2.150 1.41E+02 8.38E+01 8.13E+01 36.04 1.40E+00 D02 Unknown B7 3 28.32 4.636 4.32E+04 4.35E+04 3.33E+02 28.32 9.02E-03 D04 Unknown B7 3 28.31 4.640 4.37E+04 4.35E+04 3.33E+02 28.32 9.02E-03 Đợt 6: Định lƣợng các mẫu thí nghiệm (lần 2) Promoter 35S: PCR Standard Curve Report with PCR Amp Cycle Graph PCR Base Line Subtracted Curve Fit Data Current Date: 11-Aug-05 05:04 PM Data generated on: 03-Aug-05 at 09:21 PM. Optical data file name: Data 03-Aug-05 2121 TAQMAN DOT 2-35S-MAU.odm Plate Setup file used: Taqman 35S dot 2 3-8-05.psm Protocol file used: GMO-35S.tmo Sample volume: 25.00 ul Hot Start? No Well factor collection: Experimental Plate Comments Protocol 113 113 Cycle 1: ( 1X) Step 1: 95.0ºC for 10:00 Cycle 2: ( 45X) Step 1: 95.0ºC for 00:15 Step 2: 60.0ºC for 01:00 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC HOLD PCR Amp/Cycle Graph for FAM-490 Standard Curve Graph for FAM-490 Data Analysis Parameters Calculated threshold has been replaced by the user selected threshold 65.7. User selected baseline cycles are 2 to 20. Data analysis window is set at 95.00% of a cycle, centered at end of the cycle. Weighted Mean digital filtering has been applied. Global filtering is off. Standard Curve Spreadsheet Data for FAM-490 Units: copy number Type Identifier Rep Ct Log SQ SQ SQ Ct Ct Set SQ Mean SD Mean SD Point E02 Standard St 35S 5-8 5 26.77 3.709 5.12E+03 5.12E+03 N/A 26.77 N/A E04 Standard St 35S 5-8 6 30.13 2.806 6.40E+02 6.40E+02 N/A 30.13 N/A E06 Standard St 35S 5-8 7 32.62 1.903 8.00E+01 8.00E+01 N/A 32.62 N/A E08 Standard St 35S 5-8 8 35.24 1.000 1.00E+01 1.00E+01 N/A 35.24 N/A F02 Unknown B4 8 26.35 3.920 8.32E+03 7.88E+03 6.32E+02 26.43 1.08E-01 F03 Unknown B4 8 26.51 3.871 7.43E+03 7.88E+03 6.32E+02 26.43 1.08E-01 F04 Unknown B1 9 26.60 3.842 6.95E+03 8.16E+03 1.71E+03 26.39 2.84E-01 F05 Unknown B1 9 26.19 3.972 9.38E+03 8.16E+03 1.71E+03 26.39 2.84E-01 F06 Unknown B2 10 28.93 3.085 1.22E+03 9.45E+02 3.85E+02 29.33 5.63E-01 F07 Unknown B2 10 29.73 2.828 6.73E+02 9.45E+02 3.85E+02 29.33 5.63E-01 F08 Unknown B6 11 34.16 1.395 2.48E+01 3.33E+01 1.20E+01 33.81 4.95E-01 F09 Unknown B6 11 33.46 1.622 4.18E+01 3.33E+01 1.20E+01 33.81 4.95E-01 [...]... with PCR Amp Cycle Graph PCR Base Line Subtracted Curve Fit Data Current Date: Data generated on: 20-Aug-05 0 3:1 7 PM 03-Aug-05 at 0 9:2 1 PM Optical data file name: Plate Setup file used: Protocol file used: Data 03-Aug-05 2121 TAQMAN DOT 2 -GEN DC.odm Taqman 35S dot 2 3-8-05.psm GMO-35S.tmo Sample volume: Hot Start? Well factor collection: 25.00 ul No Experimental Plate Comments Protocol Cycle 1: ( 1X)... Sample volume: Hot Start? Well factor collection: 25.00 ul No Experimental Plate Comments Protocol Cycle 1: ( 1X) Step 1: 95.0ºC Cycle 2: ( 45X) Step 1: 95.0ºC Step 2: 60.0ºC Data collection and real-time analysis enabled Cycle 3: ( 1X) Step 1: 4.0ºC for 1 0:0 0 for 0 0:1 5 for 0 1:0 0 HOLD PCR Amp/Cycle Graph for FAM-490 114 1.04E-01 1.04E-01 9.09E-01 9.09E-01 5.83E-01 5.83E-01 115 Standard Curve Graph for...114 F10 F11 G02 G03 G04 G05 Unknown Unknown Unknown Unknown Unknown Unknown B7 B7 B10 B10 B11 B11 12 12 13 13 14 14 27.36 27.21 26.05 27.33 24.99 25.81 3.594 3.642 4.019 3.603 4.362 4.095 3.93E+03 4.39E+03 1.05E+04 4.01E+03 2.30E+04 1.25E+04 4.16E+03 4.16E+03 7.24E+03 7.24E+03 1.77E+04 1.77E+04 3.22E+02 27.29 3.22E+02 27.29 4.56E+03 26.69 4.56E+03 26.69 7.47E+03 25.40 7.47E+03 25.40 Gen tham chiếu: PCR. .. centered at end of the cycle Weighted Mean digital filtering has been applied Global filtering is off Standard Curve Spreadsheet Data for FAM-490 Units: copy number Type Identifier Rep St CR 1-4 St CR 1-4 St CR 1-4 St CR 1-4 B4 B4 B1 B1 B2 B2 B6 B6 B7 B7 B10 B10 B11 B11 1 2 3 4 1 1 2 2 3 3 4 4 5 5 6 6 7 7 Ct Log SQ SQ SQ Mean SQ SD Ct Mean 22.19 24.81 26.55 30.57 23.02 24.23 25.69 25.45 25.60 24.40 23.91... 30.59 27.44 4.612 3.709 2.806 1.903 4.268 3.861 3.371 3.452 3.403 3.806 3.970 3.965 4.426 3.575 3.630 3.631 1.726 2.785 4.10E+04 5.12E+03 6.40E+02 8.00E+01 1.85E+04 7.26E+03 2.35E+03 2.83E+03 2.53E+03 6.40E+03 9.33E+03 9.22E+03 2.67E+04 3.76E+03 4.26E+03 4.28E+03 5.32E+01 6.09E+02 4.10E+04 5.12E+03 6.40E+02 8.00E+01 1.29E+04 1.29E+04 2.59E+03 2.59E+03 4.46E+03 4.46E+03 9.27E+03 9.27E+03 1.52E+04 1.52E+04... 25.00 2.73E+03 25.00 7.56E+01 23.92 7.56E+01 23.92 1.62E+04 23.82 1.62E+04 23.82 1.07E+01 24.92 1.07E+01 24.92 3.93E+02 29.01 3.93E+02 29.01 Ct SD Point B02 B04 B06 B08 C02 C03 C04 C05 C06 C07 C08 C09 C10 C11 D02 D03 D04 D05 Standard Standard Standard Standard Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown 115 N/A N/A N/A N/A 8.57E-01 8.57E-01 . 104 104 Gen tham chiếu: PCR Standard Curve Report with PCR Amp Cycle Graph PCR Base Line Subtracted Data Current Date: 19-Aug-05 1 2:0 5 PM Data generated on: 22-Jul-05 at 0 6:1 5. Cycle 1: ( 1X) Step 1: 95.0ºC for 1 0:0 0 Cycle 2: ( 45X) Step 1: 95.0ºC for 0 0:1 5 Step 2: 60.0ºC for 0 1:0 0 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC. Step 1: 95.0ºC for 1 0:0 0 Cycle 2: ( 45X) Step 1: 95.0ºC for 0 0:1 5 Step 2: 60.0ºC for 0 1:0 0 Data collection and real-time analysis enabled. Cycle 3: ( 1X) Step 1: 4.0ºC HOLD PCR Amp/Cycle
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Xem thêm: Luận văn : BƯỚC ĐẦU XÂY DỰNG QUY TRÌNH ĐỊNH LƯỢNG CÁC SẢN PHẨM BIẾN ĐỔI GEN BẰNG PHƯƠNG PHÁP REAL-TIME PCR part 10 ppsx, Luận văn : BƯỚC ĐẦU XÂY DỰNG QUY TRÌNH ĐỊNH LƯỢNG CÁC SẢN PHẨM BIẾN ĐỔI GEN BẰNG PHƯƠNG PHÁP REAL-TIME PCR part 10 ppsx, Luận văn : BƯỚC ĐẦU XÂY DỰNG QUY TRÌNH ĐỊNH LƯỢNG CÁC SẢN PHẨM BIẾN ĐỔI GEN BẰNG PHƯƠNG PHÁP REAL-TIME PCR part 10 ppsx

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