Establishment A High Throughput Detection System Of SSR Markers Fitting For Maize Variety Based On Capillary Electrophoresis With Fluorescence

The research based on capillary electrophoresis with SSR fluorescence markers ,a high throughput detection system fitting for maize variety identification was established finally and the detection system was optimized from screening SSR markers , PCR and detection protocols and DNA extraction .The system would be applied in maize variety identification and establishing DNA fingerprinting pool of great scale maize variety. The results showed that:1. A quick DNA extraction technique which was fitting for maize variety identification and establishing DNA fingerprinting pool was explored ,and the technique was well applied in the following tissues for DNA extraction : embryo, endosperm; radicel, coleoptile , hypocotyl from germinated 1～3 days material .The extraction DNA is suitable for capillary electrophoresis with fluorescence detection system.2. As high sensitiveness of ABI3730XL DNA analyzer, common PCR amplification reaction system was not suitable for the detection system. A system fitting for PCR amplification reaction and detection was established finally through the regulation of PCR composition and cycles.3. Seven SSR primers and 192 maize hybrid were used to comparatively analyze SSR markers between capillary electrophoresis with fluorescence detection and denaturing PAGE(polyacrylamide gel electrophoresis) silver-staining detection. The results indicated that SSR markers fragment sizes explored by two detection systems were relatively similar. Capillary electrophoresis with fluorescence detection has the advantages of high accuracy, sensitiveness, cost effectiveness and high throughput. Therefore, the capillary electrophoresis with fluorescence detection system is suggested to be suitable for scanning and analyzing large-scale material. While denaturing PAGE silver-staining detection would be used in examining a few samples, especially in screening SSR markers.4.163 pairs of SSR primers selected from the internet was screened by using 15 important inbred lines widely used in production. In the 163 pairs of primers, 22 SSR pairs did not amply or "null allele", 2-8 alleles could be detected in the other 141 primers, with a total of 665 alleles, an average of 4.7 alleles per locus. A mean polymorphism information content of 0.68, with a...