SNP (Single Nucleotide Polymorphism) molecular marker related detection primer or probe for major QTL (Quantitative Trait Loci) of oil content character of cabbage type rape seeds and application of SNP molecular marker related detection primer or probe
A Brassica napus and molecular marker technology, which is applied to the determination/inspection of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., to achieve the effects of ingenious design, suitable for large-scale popularization and application, and low cost
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Embodiment 1
[0047] Example 1. Determination of the phenotype of the seed oil content trait of the Brassica napus parent and RIL population
[0048] (1) Using Brassica napus Darmor and white mustard to carry out distant hybridization, selecting the large-grain low-oil-content line GRG2462 in F2:3 as the female parent, and selecting the small-grain high-oil-content line GRD328 and GRG2462 for hybridization, The recombinant inbred lines composed of 158 lines obtained by selfing for 11 consecutive generations (Darmor of Brassica napus are from Wuhan Oil Crop Germplasm Resource Bank, and white mustard and small-grain lines GRD328 were purchased from Guizhou Hemufu Seeds Co., Ltd. Company), in Guizhou Province for many years (2012-2013 Guiyang, 2013-2014 Qinghe, 2014-2015 Changshun, 2015-2016 Guiyang, 2015-2016 Jinsha Changshun, 2016-2017 Changshun, 2017-2018 Tangtou, 2017-2018 Jinsha, 2018-2019 Changshun) seed oil content traits were investigated.
[0049] (2) Using live seedlings, row spacin...
Embodiment 2
[0052] Example 2, the acquisition of high-quality SNP data set of RIL population
[0053] The CTAB method was used to extract the total leaf DNA, and the total leaf DNA of each material of the RIL population was extracted. The specific method is as follows:
[0054] Rinse the young leaves in 10% ethanol; then cut 0.1-0.2 g of leaves into a grinding bowl, quickly grind with liquid nitrogen to powder, and put them into a 2 mL centrifuge tube; add 700 μL of preheated DNA extraction solution; After mixing, put it in a 65°C water bath for 1 hour, and mix once every 10-15 minutes; add 700 μL of mixed solution (phenol:chloroform:isoamyl alcohol=25:24:1), gently invert and mix for 10 minutes; at room temperature, Centrifuge at 10 000 × g for 15 min; pipette the supernatant into a new 2 mL centrifuge tube; add an equal volume of mixed solution (chloroform: isoamyl alcohol = 24: 1), invert and mix, let stand for 5 min, 10000 × g, centrifuge for 15 min , suck the supernatant into a new ...
Embodiment 3
[0056] Embodiment 3, the construction of high density genetic map
[0057] According to the above Example 2, a high-quality population SNP dataset was obtained and constructed using MadMapper (http: / / cgpdb.ucdavis.edu / XLinkage / MadMapper / ) and HighMap software (http: / / highmap.biomarker.com.cn / ). High-density SNP genetic linkage map of the RIL population. In order to improve the speed of operation, firstly, use the RECBIT program in MadMapper software to remove redundant SNPs (two or more SNPs show co-segregation in the population), data missing ratio ≥ 25% of SNPs in the 158 lines, and SNPs in the population. SNPs with an allelic segregation ratio figure 2 shown.
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