Specific sequence characterized amplified region (SCAR) marker for gametocidal chromosome 2C of aegilops
A technology for chromosomes and gametes, which is applied in the field of gameticidal chromosome 2C-specific SCAR markers of Capt.
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Embodiment 1
[0033] Example 1. Acquisition of Specific SCAR Marker on Chromosome 2C of Capeatia genus Gametocidal
[0034] 1. Screening and cloning of the gametocidal chromosome 2C-specific RAPD marker OPF03 marker of the genus Goatia
[0035] Using 40 RAPD primers, amplified and screened common wheat "China Spring", common wheat "Chinese Spring"-gametocidal chromosome 2C disomy addition line, and Goatia stigmatis, and isolated and cloned 2C genome-specific amplified fragments .
[0036] Among them, the 20μl system amplified by PCR
[0037] Template DNA 50ng
[0038] 10xPCR buffer 2.5μl
[0039] 1.5mM MgCl 2 2.5μl
[0040] dNTP Mix (2.5mM) 1.5μl
[0041] Random Primer (10μM) 1μl
[0042] r Taq polymerase (5U / μl) 0.25μl
[0043] Distilled sterile water up to 20μl
[0044] 10x PCR buffer was purchased from Treasure Bioengineering (Dalian) Co., Ltd., catalog number R10T1.
[0045] PCR program: pre-denaturation at 94°C for 3 min, denaturation at 94°C for 15 s, anneali...
Embodiment 2
[0068] Example 2, the application of the gametocidal chromosome 2C-specific SCAR marker of the genus Goatia
[0069] with SCAR primers 2C-F 586 , 2C-R 586 F 1 and F 1 F 2 Generation was identified. The common wheat "China Spring" was used as the control.
[0070] The result is as image 3 As shown, 1. is the common wheat "China Spring"; 2-12. is the F obtained by crossing the "China Spring"-Thinopyrum elongatum 7E disomy addition line and "China Spring"-gametocidal chromosome 2C disomy addition line 1 generation; 13. is DL2000Marker; 14~25. is F 1 F 2 generation.
[0071] It can be seen from the figure that in F 1 A product of 586 bp was amplified in all the materials of the first generation, which was in line with theoretical expectations. After sequencing, the fragment had the sequence table sequence 2 from the 5' end of the 405th to 990th nucleotide, which is unique to the gametocidal chromosome 2C, F 1 All generations showed positive results.
[0072] while in ...
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