Establishment Of InDel Multiplex Detection System Suitable For Identification Of Maize Varieties

Our country is a large agricultural country,paying attention to agricultural production.Maize is one of the most important forage crops,the planting area is wide,and the variety of maize is increasing year by year.So,it is very important to evaluate the authenticity and purity of Maize Varieties.In recent years,the rapid development of molecular marker technology,people are eager to apply the latest research results to the daily work.In order to apply the In Del markers and multiplex PCR technology to the test of maize varieties,the core primer combination method was used to construct 20 PCR combinations on the basis of the 10 pairs of core primers,40 fluorescence capillary electrophoresis.In the first part of this study,In order to improve the detection efficiency with molecular marker,the multiple PCR detection system was constructed.In this study,10 major materials were used to evaluate the single pair PCR with 238 pairs of In Del primers.According to the software quality evaluation results,product range,and the principle of chromosome uniform distribution,30 pairs of primers were selected from192 primers with better performance to form three groups of core primer combinations with amplified products in the range of 80-200 bp and 200-400 bp,There were 10 pairs of primers distributing in different chromosomes for each primer combination.Based on core primer combination and comprehensive consideration on chromosome distribution,base fragment and primers fluorescent color,we established two groups of corn test 20 PCR system and a group of 40 fluorescent capillary electrophoresis.The second part is the identification of the purity of the particular maize variety Jing ke968.For different samples,40 pairs of primer combinations were needed to identify,but for a specific variety of corn,we can continue to select the best combination of purity identification.This combination is suitable for a specific variety and the combination of high stability,for different quality of DNA are applicable.In this study,7 pairs of primers were selected from the best combination of the 40 pairs of primers.The optimal combination of these two pairs of primers was selected from the 968 pairs of primers.Compared with the 40 pairs of primers,the results showed that these 7 pairs of primers can not only play the role of purity identification,but also save some experimental reagents.