KASP molecular marker of wheat powdery mildew resistance gene Pm21 derived from haynaldia villosa and application of KASP molecular marker
A powdery mildew resistance gene and molecular marker technology, which is applied in the field of biotechnology and genetic breeding, can solve the problems affecting the efficient utilization of Pm21 gene, and achieve the effects of accelerating the utilization of high-efficiency breeding, high-throughput detection and tracking, and improving the utilization of transgenics
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[0029] Example 1 Wheat Powdery Mildew Resistance Gene Pm21 The acquisition and application of KASP molecular marker KASP-Pm21
[0030] 1. Preparation of DNA amplification template
[0031] wheat powdery mildew resistance Pm21 The vector of the gene is the vector constructed in He et al. (2018), which is ready for use. The rest of the DNA amplification templates were extracted by CTAB method from 1 T. villosa and 2 Pm21 Transgenic lines, 8 carrying Pm21 6VS translocation lines and 20 do not carry Pm21 The genomic DNA of the one-leaf stage leaves of common wheat, the specific operation steps are as follows:
[0032] (1) Cut fresh leaves and freeze and grind them with liquid nitrogen. Take 0.5g of ground powder for each sample in a 2mL centrifuge tube;
[0033] (2) Add 600 μL of 65°C preheated CTAB extract to each sample, and bathe in 65°C water bath for 45 minutes;
[0034] (3) Cool the centrifuge tube to room temperature, add the same amount of chloroform-isoamyl alcohol...
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