适于多物种的通用尾巴序列设计及通用体系的建立OA北大核心CSTPCD

[Objective]Fluorescent capillary electrophoresis,for its high detection throughput and resolution,was in extensive applications in various scenarios,including individual identification,variety discrimination,and species identification.The tailed-sequence offers an efficient solution for the extensive application of the fluorescent capillary electrophoresis platforms.To address the limitation of existing tailed-sequences in meeting the demands of multi-species applications,this study developed an efficient Universal Tailed-Sequence(UTS)design tool based on encoding translation technology.Utilizing this tool,UTS were designed to construct a versatile PCR system and protocol suitable for multiple crops,thereby enhancing the throughput and flexibility of fluorescent electrophoresis.[Method]An efficient UTS design tool was designed based on encoding translation technology and 3 755 commonly used Chinese characters were encoding translated with filtering criteria including GC content,hairpin,and the self-dimer annealing temperature.The BLAST tool was used to assess the homology of UTS generated by the design tool on various biological genomes.The UTS was experimentally evaluated on crops such as maize,tomatoes,peppers,and watermelons,constructing a species-universal experimental system and protocol.[Result]The design tool,through encoding translation and screening,generated a total of 7 436 833 high-quality candidate UTS,accounting for 52.74%of all character combinations.Six selected UTS were demonstrated better specificity across 20 crop genomes by BLAST results compared to M13.With optimization of the general primer amplification procedure,the success rate of general primer amplification across multiple species reached or exceeded 95%,showing strong stability.[Conclusion]This study utilized encoding translation technology to develop UTS and paired them with a species-universal amplification system and protocol,which may provide a high-throughput,cost-effective universal detection method for fluorescent electrophoresis.