Transgenic Quantitative Detection Method Based on PCR-free High-throughput Sequencing Technology
Quantitative GMO detection is essential for ensuring food safety and protecting consumer rights to information.Al-though digital PCR is currently considered the gold standard for accurate nucleic acid quantification,the lack of validated new quantification technologies limits its applications.However,the emergence of high-throughput sequencing has opened up new possibilities for solving this challenge.While high-throughput sequencing is primarily used for qualitative nucleic acid sequence determination,its potential for quantitative analysis has yet to be fully explored.In this study,plasmid DNA standard materials containing transgenic T-NOS,P-35S,CP4-EPSPS,and soybean housekeeping gene Lectin were used as the detection objects.The method of library construction without amplification was adopted to compare the differences between NGS,third-generation sequencing,and digital PCR quantification.The results showed significant differences between NGS sequencing results and digital PCR results,highlighting the challenges and demands in current nucleic acid quantitative analysis technologies.However,it is worth noting that the results of third-generation sequencing were consistent with the digital PCR detection results,demon-strating its potential as a precise method for quantifying transgenic nucleic acids.
万方数据
维普
