A group of rice ssr molecular markers suitable for capillary electrophoresis detection technology and its application
A technology of capillary electrophoresis and molecular markers, which is applied in the field of crop molecular biology, can solve the problems that different batches of reaction data are difficult to be processed uniformly, different allelic variations are difficult to accurately identify, non-automation, etc., to achieve accurate fragment size and calculate fragment size , the effect of improving efficiency
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Embodiment 1
[0032] Example 1 Determination of rice SSR molecular markers and primers
[0033] 36 rice varieties were selected for SSR primer screening. Using denaturing polyacrylamide gel electrophoresis, 800 pairs of SSR primers (http: / / www.gramene.org / ) were used to select primers with amplified sites, clear band patterns, and high polymorphism as optional primers. According to the molecular weight range and PIC value of the alleles of the optional primers in the 36 varieties, that is, each combination is composed of as many primers as possible, and the allele ranges of all primers in the group do not overlap, four primer sets are formed and combined. The primers with FAM, VIC, NED and PET fluorescent groups were synthesized respectively. See Table 1. Those skilled in the art can understand that other fluorophores including FAM, VIC, NED, PET can also be selected in the art to modify the four sets of primers in Table 1, as long as the primers in each group are labeled with the same fl...
Embodiment 2
[0037] Embodiment 2 uses the SSR molecular marker provided by the present invention to distinguish rice varieties
[0038] (1) Rapid extraction of DNA
[0039] DNA was extracted from rice seeds by alkaline boiling method. This method is quick and easy to operate, has no toxic and harmful reagents, is suitable for DNA preparation in the field of plant molecular biology, and 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 rice seeds and place them in a 1.5mL centrifuge tube; add 400 μL NaOH (1M) to the centrifuge tube to ensure that the seeds are completely soaked, and bathe in boiling water for 5 min; add 200 μL Tris-HCl ( 1M, pH 8.0), in a boiling water bath for 1 min; add 200 μL TE buffer (pH 8.0), fully dissolve and set aside.
[0040] (2) Quality and quantity of DNA samples
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