Hybrid Chinese tuliptree rapid molecular detection specific primer and use method thereof
A technology of hybrid Liriodendron and molecular detection, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems that cannot achieve rapid and accurate identification, affect the reliability of detection results, and RAPD Large impact and other problems, to achieve the effect of simple and fast operation, convenient material collection and high sensitivity
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Embodiment 1
[0033] Example 1: Primer LT008L / R was used for species identification of Liriodendron tulipifera, Liriodendron tulipifera and hybrid Liriodendron tulipifera.
[0034] The primer pair LT008L / R was used to identify 15 samples each of Liriodendron tulipifera, Liriodendron tulipifera and Hybrid tulipwood. Results The 190bp product was specifically amplified in the Liriodendron tulipifera population, the 180bp product was specifically amplified in the Liriodendron tulipifera population, and both 180bp and 190bp products were amplified in the hybrid Liriodendron tulipifera population. 10 μL of the reaction system for PCR, including 1 μL of template DNA, 1 μL of 10×Buffer, 0.2 μL of 10 mM dNTPs (Shanghai Jierui Biological Co., Ltd.), 25 mM MgCl 2 1 μL, F-primer 0.25 μL, R-primer 0.25 μL, Ampli TaqGold 0.05 μL (Shanghai Jierui Biological Co., Ltd.), ddH2O 6.25 μL. The PCR reaction program is: pre-denaturation at 94°C for 4 minutes; gradient cooling amplification: 15 cycles of 94°C fo...
Embodiment 2
[0038] Example 2: The primers LT008L / R were used to identify the species of Liriodendron tulipifera, Liriodendron tulipifera and Hybrid tulipwood different from Example 1.
[0039] The primer pair LT008L / R was used to identify 15 samples of Liriodendron tulipifolia, Liriodendron tulipifera and hybrid Liriodendron tulipifera different from those in Example 1. Results The 190bp product was specifically amplified in the Liriodendron tulipifera population, the 180bp product was specifically amplified in the Liriodendron tulipifera population, and both 180bp and 190bp products were amplified in the hybrid Liriodendron tulipifera population. kind of product. The PCR prescription system and PCR reaction procedure are all the same as the detection and analysis steps of Liriodendron tulipifera in Example 1. The details of the samples used are shown in Table 2, and the test results are shown in figure 2
[0040] Table 2. The specific amplification results of 15 samples of Liriodendr...
Embodiment 3
[0042] Embodiment 3: Using the specific primer LT008L / R of the present invention to control the pollination progeny identification of Liriodendron tulipifera multi-paternal mixing, using the primer pair LT008L / R, the female parents are Nanka, Lewis and Lushan, and the range of the male parents is known ( Lushan, Fuyang, Missouri, South Carolina, Lewis) 86 samples of the control pollination progeny for species identification. The PCR prescription system and PCR reaction procedure are all the same as the detection and analysis steps of Liriodendron tulipifera in Example 1. The specific results of the experiment are shown in Table 3.
[0043] Table 3. The identification results of the primer LT008L / R on the pollination progeny of Liriodendron tulipiflora mixed with multiple male parents
[0044]
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