A rapid detection kit for simultaneous detection of multiple transgenic rapeseed lines and its application
A kit and transgenic technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of high detection cost, unsuitable multi-strain screening and detection of genetically modified products, environmental pollution, etc. Achieving good stability and suitable for commercial promotion
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Embodiment 1
[0032] Embodiment 1: PCR detection method and sensitivity, specificity experiment
[0033] (1) Materials and methods
[0034]Transgenic standard products were purchased from American Standard Materials AOCS or European Standard Materials ERM, including: transgenic rapeseed Topas19 / 2 (AOCS0711-D4), transgenic rapeseed RF2 (AOCS0711-C3) purchased from American Oil Chemists' Society (AOCS, Urbana, USA) ), transgenic rapeseed Ms1 (AOCS0711-A3), genetically modified rapeseed Ms8 (AOCS0306-F7), genetically modified rapeseed Rf3 (AOCS0306-G6), transgenic rapeseed T45 (AOCS0208-A6), genetically modified rapeseed Rf1 (AOCS0711-B2), transgenic rapeseed GT73 (AOCS0304-B2), transgenic rape MON88302 (AOCS1011-A), non-transgenic rape (AOCS 0304-A2), transgenic soybean MON89788 (AOCS 0906-B2), non-transgenic soybean (AOCS 0906-A), transgenic corn GA21 ( AOCS0407-B), non-transgenic maize (AOCS0411-CD) and transgenic rapeseed 73496 (ERM-BF434e) purchased from European Reference Materials (ERM...
Embodiment 2
[0054] Example 2: Verification of microfluidic chip stability
[0055] The stability of the microfluidic chip of the present invention is verified, and the microfluidic chip that is not added with taurine and freeze-dried is used as a control to verify the conditions of the chip at room temperature (25°C), -20°C and repeated freezing and thawing Under the chip detection performance.
[0056] The detection chips for CRUA, transgenic rapeseed RF2, and transgenic rapeseed T45 were placed at room temperature (25°C) and -20°C for 3 months, 6 months, and 12 months, respectively, to test the stability of the kit. The verification experiment was carried out with transgenic rapeseed Topas19 / 2 (AOCS0711-D4) at a concentration of 5ng / uL. The experimental results are shown in Table 3 below. When the microfluidic chip of the present invention is placed at room temperature (25°C) for 3 months, 6 months, and 12 months, there is no significant difference in the Ct value and Tm value, and the...
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