SNP molecular marker for identifying and controlling purple character formation of Chinese cabbages and application thereof
A technology of DNA molecules and purple, which is applied in recombinant DNA technology, microbe determination/inspection, DNA/RNA fragments, etc., and can solve the problem of rare research on key genes for purple formation
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Embodiment 1
[0079] Example 1, the acquisition of molecular markers
[0080] In order to screen and obtain purple Chinese cabbage resources better and faster, and to promote the practice of molecular marker-assisted selection breeding, a purple Chinese cabbage material (09N-742) and a green Chinese cabbage material (09-680) were regenerated Sequencing data analysis found that there is a T / A mutation SNP site at base 31593020 of chromosome A03 ((300th nucleotide in Sequence 4 of the Sequence Listing)), and the SNP site is named A0331593020T / A position point. According to the A0331593020T / A site, design the following specific primers that can be used in KASP technology as molecular markers: upstream primer A0331593020T / A-FF, upstream primer A0331593020T / A-FV and downstream primer A0331593020T / A-R.
[0081] The above-mentioned A0331593020T / A-FF, A0331593020T / A-FV and A0331593020T / A-R primers were entrusted to UK LGC (Laboratory of the Government Chemist Government Chemist Laboratory) Limited...
Embodiment 2
[0090] Example 2, Application of A0331593020T / A site in identified purple and green materials
[0091] 1. DNA extraction
[0092] Genomic DNA of 518 kinds of cabbage materials to be tested in Table 1 were extracted by conventional CTAB method.
[0093] 518 materials are hybrid offspring BC of parents 09N-742 and 09-680 6 f 2 The specific methods are as follows: cross the purple parent material 09N-742 and the green parent material 09-680 to obtain F1; then use the green parent material 09-680 as the backcross parent for 6 generations to obtain a F1 consisting of 518 individual plants BC 6 f 2 (group.
[0094] The quality of the extracted DNA was tested by agarose electrophoresis and Nanodrop2100 respectively, and it was found that the extracted genomic DNA met the relevant quality requirements, that is, agarose electrophoresis showed a single DNA band without obvious dispersion; Nanodrop2100 detected A260 / 280 between 1.8-2.0 between (DNA samples without protein contamina...
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