A Chinese cabbage burr indel marker and its detection primer and application
A technology for detecting primers and Chinese cabbage, which is applied in the field of molecular biology and can solve problems such as lack of markers and hindering breeding.
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Embodiment 1
[0039] Example 1. Development of InDel labeled primers
[0040] Taking Chinese cabbage burr formation-related genes as candidate genes, hairy (G291) and hairless (ZHB) Chinese cabbage genome-wide resequencing was performed using resequencing technology. According to the resequencing results, the candidate genes in hairy and hairless Chinese cabbage were compared, and primers were designed for one of the InDel marker TR-1. The sequence comparison of the Chinese cabbage InDel marker TR-1 is shown in figure 1 shown. The area marked by a line in the figure is the InDel marker TR-1.
[0041] Hairy Chinese cabbage (G291) has a 12 bp DNA insertion compared to hairless Chinese cabbage (ZHB). The detection primers for Chinese cabbage InDel-labeled TR-1 were designed, and the primer sequences are shown in SEQ ID NO.1 and SEQ ID NO.2.
Embodiment 2
[0042]Application of embodiment 2.InDel labeled primers
[0043] 2.1 Extraction of plant material and template DNA
[0044] Select the first generation of Chinese cabbage hybrid commercial varieties Qingyan Chunbai 2, Degao Baihe, Xibai 45, Xibai 8, Zhonghua 1, Qingyan CR20, Qingyan Xiabai 2, Zhonghua 1, Gaokang 2 , Xibai 5, Xiayou 2, Jintongyunv, Gongguan 2, Fengkang 60, Huayangbai, Degaoxikang 65 and Chinese cabbage inbred lines Guangdong Zao, Y2, L41, L46, AM4-1, AM3 -1, AM5-2, AM9-1, AM94, AM30, AM54, AM72-1, AM10-1, AM56-1, and hybrid F2 leaves of ZHB and G291 were used as materials.
[0045] The total DNA was extracted by the modified CTAB method, and the specific steps were as follows:
[0046] 1. Take 1-10g of fresh Chinese cabbage leaves and quickly grind them into powder in liquid nitrogen;
[0047] 2. Transfer the frozen powder into a pre-cooled centrifuge tube, add an equal volume (w / v, g / ml) of 2×CTAB extraction buffer, and incubate at 65°C for 20 minutes;
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