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用L25(56)正交设计建立和优化糯玉米SSR-PCR反应体系;利用370对SSR引物对糯玉米两亲本及F1进行扩增,筛选抗丝黑穗病的相关引物。结果表明:20μL糯玉米SSR-PCR最优反应体系为1.5 U DNA聚合酶,1.0 nmol/μL镁离子,80 ng模板DNA,0.1 pmol/μL引物,0.15μmol/μL d NTPs,1倍扩增缓冲液;370对SSR引物对两亲本及F1进行扩增的条带显示,有差异且条带清晰的引物为87对。确立了糯玉米SSR-PCR最优反应体系,筛选并得到了87对多态性引物。
The orthogonal design of L25 (56) was used to establish and optimize the SSR-PCR reaction system of waxy maize. 370 pairs of SSR primers were used to amplify the waxy maize parents and F1, and the relative primers were screened for resistance to head smut. The results showed that the optimal SSR-PCR reaction system for 20 μL waxy corn was 1.5 U DNA polymerase, 1.0 nmol / μL magnesium ion, 80 ng template DNA, 0.1 pmol / μL primer, 0.15 μmol / μL dNTPs, Fluids; 370 pairs of SSR primer pairs amplicons and F1 amplified bands showed that there are 87 pairs of primers with differential and clear bands. The optimal SSR-PCR reaction system for waxy maize was established, and 87 pairs of polymorphic primers were screened out.