Method of acquiring rDNA ITS2 sequence for identification of dendrobium candidum
A Tiepifengdou and sequence technology, which is applied in the field of identification of medicinal materials source varieties, can solve the problems of affecting DNA extraction efficiency, sample DNA degradation, low amplification efficiency, etc., and achieve the effects of easy control, high accuracy and simple operation.
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Embodiment 1
[0022] Identification of Example 1 Tiepi Fengdou
[0023] 1. Experimental materials
[0024] In this case, a total of 69 samples of Tiepi Fengdou were collected, and the specific information is shown in Table 1.
[0025] Table 1 Tiepi Fengdou sample information
[0026]
[0027]
[0028]
[0029] 2. Experimental equipment
[0030] Desktop high-speed centrifuge, PTC-100 gene amplification instrument, electrophoresis instrument, ultraviolet gel imaging analysis system, MM400 ball mill (Germany Retsch).
[0031] 3. DNA extraction
[0032] Take 20-30 mg of dried Tiepi Fengdou medicinal materials, chop them up and crush them with MM400 ball mill, add 500-1000 μL of nuclear separation solution, mix well, centrifuge at 7000 r / min for 5 minutes, discard the supernatant, add 500-1000 μL of nuclear separation solution, repeat This step is repeated 3-5 times until the supernatant is not viscous. The remaining steps were carried out according to the standard operating method...
Embodiment 2
[0050] The identification of embodiment 2 Dendrobium officinale base plant
[0051] Basically the same as Example 1, except that the original plant of Dendrobium candidum was used as the sample, DNA was extracted according to the above method, and the ITS2 sequence was amplified. As a result, the target sequence was also successfully obtained and the identification was completed.
Embodiment 3
[0052] The identification of embodiment 3 dendrobium officinale seed seedlings
[0053] Basically the same as Example 1, the difference is that Dendrobium officinale seeds and seedlings are used as samples, DNA is extracted according to the above method, and the ITS2 sequence is amplified. As a result, the target sequence can also be successfully obtained and the identification is completed.
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