Kelp gametophyte DNA solution preparation method and sex determination method
A gametophyte and kelp technology, applied in the field of marine biotechnology and seaweed molecular genetics, can solve the problems of negative impact of the environment and experimenters, cumbersome experimental steps, labor and financial resources, etc., to save sample storage space, reduce costs and errors The effect of low ratio
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Embodiment 1
[0041] Example 1: To verify the feasibility of the preparation method of Laminaria gametophyte DNA solution which can be used for PCR amplification.
[0042] 6 kelp species 901 kelp (901), Zaohoucheng kelp (ZHC), Benniu kelp (BN), Dongfang No. 6 kelp (Dong 6) provided by the Germplasm Preservation Laboratory of Seaweed Center of Shandong Oriental Ocean Technology Co., Ltd. , Korean kelp (HG), Lianza kelp (LZ), 3 gametophytes for each male and female, a total of 36 gametophytes (see Table 1 for the number of gametophytes). Utilize two methods to obtain the laminar gametophyte genomic DNA solution: one utilizes the plant genomic DNA extraction kit of Tiangen Company to extract 36 laminaria gametophyte DNAs; Laminaria gametocytes with a wet weight of about 15-30 mg were placed in a 1.5 mL sterilized centrifuge tube, and 500 μL of sterilized deionized water was added, thoroughly ground, boiled in water for 20 minutes, and allowed to settle at room temperature for 30 minutes) to ob...
Embodiment 2
[0045] Example 2: Verification of the correctness of double-primer PCR sex identification of Laminaria gametophytes.
[0046] 631301♀♂ mixed gametophyte clone provided by the germplasm preservation laboratory of Muping Branch of Seaweed Center of Shandong Oriental Ocean Technology Co., Ltd., S.ang 1301 ♀♂ mixed gamete clone, S.jap 1301♀♂ mixed gametophyte clone, 491302♀♂♂ mixed, 261102♀♂ mixed; 5 Laminaria female gametophytes and 5 Laminaria male gametophytes were randomly selected from the Laminaria gametophyte sample in Example 1, and a total of 15 clonal lines were tested (gametophyte numbering See Table 2). Take an appropriate amount of gametophyte clones in a 1.5 mL sterilized centrifuge tube, add 500 μL of sterilized deionized water, grind thoroughly, bathe in boiling water for 20 minutes, and settle at room temperature for 30 minutes to obtain a DNA solution of 15 Laminaria gametophyte clones. The first method is to use Laminaria gametophyte female-specific primer H...
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