Traditional Chinese medicine alpinia oxyphylla EST-SSR marker primer and application thereof
A technology for labeling primers and traditional Chinese medicines, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve the problems of the diversity and content differences of the medicinal ingredients of the nutraceutical fruit
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[0030] Test material:
[0031] Table 1 Information of puzzle samples in different regions
[0032]
[0033]
[0034] 1. Select the tender leaves of 17 regions to extract total DNA, and refer to the extraction method. Steps in the instructions of the plant genomic DNA extraction kit.
[0035] 2, utilize the EST-SSR marker designed by the present invention to analyze, and the concrete steps are as follows:
[0036] 2.1 EST-SSR primer screening and amplification:
[0037]2.1.1 PCR reaction system: The optimal reaction system for this experiment is 20 μl: 10 μl 2×Power Taq PCRMasterMix (ie 10 μl DNA Polymerase, Buffer containing Mg 2+ , 2.0mM / μl dDTP Mixture), 2μl DNA, 2μl primer (concentration 10μmol / L), 6μl ddH 2 O (sterilization).
[0038] 2.1.2 PCR reaction program: pre-denaturation at 94°C for 2min, denaturation at 95°C for 45sec, annealing for 45sec (annealing temperature is the optimum temperature for each primer), extension at 72°C for 10min, 36 cycles, and sto...
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