Molecular identification method of high-digestibility sorghum strain
A Molecular Identification, High Digestibility Technology
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Embodiment 1
[0016] Example 1: Molecular marker identification of highly digestible sorghum lines
[0017] 1. Crossing of high digestibility lines: Brown midrib sorghum line U68 (type bmr-6) was used to cross with sorghum line Tx623B.
[0018] 2. The first backcross: backcross with the hybrid as the female parent and sorghum Tx623B as the male parent to obtain BC 1 f 1 .
[0019] 3. Molecular marker identification of offspring individual plants: BC 1 f 1 20 plants were planted, DNA was extracted from individual plants, and the individual plants carrying the brown midrib gene were identified by PCR amplification.
[0020] The identification procedure is as follows:
[0021] ①. Use primer B-1 (pre-primer: GACGGCCGGCACGCG; post-primer: TTCAGACTTCAGACCAAC) to amplify the DNA of these 20 individual plants (total system: 30 μL: DNA 100ng 2mM primer 5 μL, 2 Mm dNTP 6 μL, KOD enzyme 0.5 U, 15 μL of 2-fold buffer, and make up the rest with double-distilled water. The amplification program is:...
Embodiment 2
[0026] Example 2: Molecular marker identification of high digestibility sudan grass strains
[0027] 1. Hybridization of high digestibility strains: Brown midrib sorghum line U68 (type bmr-6) was used to cross with sudangrass line S722.
[0028] 2. The first backcross: backcross with the hybrid as the female parent and Sudan grass line S722 as the male parent to obtain BC 1 f 1 .
[0029] 3. Molecular marker identification of offspring individual plants: BC 1 f 1 20 plants were planted, DNA was extracted from individual plants, and the individual plants carrying the brown midrib gene were identified by PCR amplification.
[0030] The identification procedure is as follows:
[0031] ①. Use primer B-1 (pre-primer: GACGGCCGGCACGCG; post-primer: TTCAGACTTCAGACCAAC) to amplify the DNA of these 20 individual plants (total system: 30 μL: DNA 100ng 2mM primer 5 μL, 2 Mm dNTP 6 μL, KOD enzyme 0.5 U, 15 μL of 2-fold buffer, and make up the rest with double-distilled water. The amp...
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