InDel mark tightly linked with wild peach germplasm anti-green-peach-aphid properties and application thereof
A pp-indel-23, marker technology, applied in the field of molecular breeding, can solve problems such as poor pesticide effect and aphid variation, and achieve the effect of saving land
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[0019] Example 1: Screening of aphid-resistant plants from the offspring population of peach germplasm 2013-04-20R
[0020] 1. Collect individual leaves from 2013-04-20R offspring, and extract leaf DNA with a plant DNA extraction kit; dilute the DNA to 20ng / ul;
[0021] 2. Select the detection site molecular marker primers Pp-InDel-23F and Pp-InDel-23R in the invention, and amplify the molecular markers by PCR. The PCR reaction system is: total volume 20μl, 1×PCR buffer, 1.5mmol / L MgCl2, 0.2mmol / LdNTPS, 0.4mmol / L primers Pp-InDel-23F and Pp-InDel-23R, 25ng template DNA, 1U Tag enzyme ;PCR reaction conditions: Denaturation at 95°C for 3min, followed by 35 cycles at 94°C for 30s, 53°C for 25s, 72°C for 15s, and finally extension at 72°C for 5min.
[0022] 3. After the amplified product is separated by electrophoresis on a 6% polyacrylamide gel, detect the band type to see if it is a heterozygous band type or contains a 93bp resistant band (attached figure 1 ). The 6% polyacr...
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