SSR molecular markers for identifying cannabis sativa varieties and application of SSR molecular markers
A technology of molecular markers and marijuana, applied in the field of molecular breeding and identification of crops, to achieve the effect of accurate and reliable results, accurate results and improved efficiency
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Embodiment 1
[0049] Embodiment 1 Determination of cannabis SSR molecular markers and primers
[0050] According to 22 cannabis varieties (Yousha-31, Longma No. 1, Longmai No. 2, Huoma No. 1, Qingmai No. 1, Longmai No. 3, Longma No. 1, Hanma No. 2, Hanma No. 1 , Longma No. 2, Grecia, Hanma No. 5, Hanma No. 4, Muma No. 1, Qingma No. 2, Xianma No. 1, Longma No. 4, Qingmai No. 3, Qingmai No. 4, The gene sequences of Hanma No. 6, Hanma No. 7, and Longmai No. 5) were searched for SSR sites, and 300 pairs of SSR primers were designed; Grecia, Huoma No. 1, Longmai No. 5, and Hanma No. 7 were extracted , Longmai No. 1 and Yousha-31 DNA, respectively use these 300 pairs of primers to carry out PCR and polyacrylamide gel electrophoresis for these six varieties, analyze and compare the annealing temperature, whether it can be amplified, and the band of each pair of primers Parameters such as specificity, clarity, and whether a stable single band can be obtained in the six varieties, and 210 pairs of ...
Embodiment 2
[0053] Embodiment 2 Utilizes the SSR molecular marker provided by the present invention to distinguish cannabis varieties
[0054] (1) Rapid extraction of DNA
[0055] DNA extraction from hemp seeds was carried out by CTAB method. This method is quick and easy to operate, and the quality of the extracted DNA is high. It is suitable for DNA preparation in the field of plant molecular biology, and it is of great significance for greatly shortening the time of seed purity inspection and transgenic detection, improving detection efficiency, and reducing detection costs. The specific operation steps are as follows: take a few hemp seeds and grind them into powder, put them in a 1.5mL centrifuge tube; add 700 μL of CTAB extract to the centrifuge tube, and incubate at 65°C for 30 minutes; add 500 μL of chloroform-isoamyl alcohol ( 24:1), shake vigorously, centrifuge at 12000rpm for 10min; take the supernatant, add 0.7 times the volume of isopropanol, and centrifuge at 12000rpm for 5...
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