Detection primer, detection kit and detection method of dehiscence resistance of Sesamum indicum
A detection method and a technology for detecting primers, which are applied in the field of molecular biology, can solve the problems of cumbersome detection and low repetition rate of sesame, and achieve the effects of improving breeding efficiency, accurate detection methods, and shortening the breeding cycle
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Embodiment 1
[0035] Acquisition of primers for detection of sesame resistance to dehiscence:
[0036] 1. Obtaining Zhengzhi InD01: using the anti-capsulation material "UCR82-15NS" (provided by D.M.Yermanos, Riverside, USA in 1983) as the female parent and Yuzhi No. 11 as the male parent, F 1 , selfing to obtain F 2 ; select F 2 A single plant of Zhongkang Xijiao was backcrossed with Yuzhi No. 11 to obtain BC 1 f 1 , selfing to get BC 1 f 2 ; select BC 1 f 2 A single plant of Zhongkang Xijiao was backcrossed with Yuzhi No. 11 to obtain BC 2 f 1 , selfing to get BC 2 f 2 ; select BC 2 f 2 A single plant of Zhongkang Xijiao was backcrossed with Yuzhi No. 11 to obtain BC 3 f1 , selfing to get BC 3 f 2 ; 2 generations of self-crossing, get BC 3 f 4 , the material with good capsule resistance, good capsule formation and disease resistance was selected and named Zhengzhi InD01.
[0037] Zhengzhi InD01, a sesame anti-capsulation-resistant material, Yuzhi No. 11, and its F2 segreg...
Embodiment 2
[0052] Sesame anti-capsulation detection kit, including: 10 μmol / μL 2 mL of the forward primer of Example 1, 10 μmol / μL of 2 mL of the reverse primer of Example 1, 1 mL of 5 U / μL Taq enzyme, 10 mL of 10×PCR buffer, 10 mM 2mL of dNTPs / μL, ddH 2 O 10mL.
Embodiment 3
[0054] Sesame anti-cracking capsule detection method, comprises the steps:
[0055] 1) Using the CTAB method to extract F from Zhengzhi InD01 and other sesame materials 2 The DNA of the leaf tissue of the progeny plants of the above-generation groups;
[0056] 2) PCR reaction system: 10 μL, containing 1 μL template DNA (25-50ng / μL), 0.1 μL Taq enzyme (5U / μL), 1 μL 10×PCR buffer, 0.2 μL dNTPs (10mM / μL), 0.2 μL of forward primer (10 μM / μL), 0.2 μL of reverse primer (10 μM / μL), 7.3 μL of ddH 2 O; Amplification program: pre-denaturation at 94°C for 1min; denaturation at 94°C for 30s, annealing at 57°C for 30s, extension at 72°C for 30s, 35 cycles, extension at 72°C for 10min;
[0057] 3) The amplified product was separated by 9% non-denaturing polyacrylamide gel electrophoresis, the electrophoresis buffer was 0.5×TBE, and the electrophoresis was separated at 150V constant power for 1h10min. After electrophoresis, the color was developed by silver staining, and the band pattern w...
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