PCR产物的高通量测序方法及优化

Methods and Optimization of High Throughput Sequencing Technologies on PCR Products

PCR产物的高通量测序被广泛应用于功能基因筛选、肿瘤相关基因的突变和甲基化检测等。在高通量测序技术中,建库和上机测序实验一直是决定最终DNA数据质量的关键。作者优化了PCR产物样品的DNA文库制备条件和体系,设计了适合PCR产物特点上机测序方法,将已建库的PCR样品中混入一定量的基因组标准品后再上机测序,由此保证了测序数据的高质量和低冗余度。为低多样性DNA样品高通量测序技术方法运用提供方法上的参考。

The development of the new generation sequencing technology extends the application field of gene sequencing. High throughput sequencing of PCR products has been widely used in functional gene screening,mutation and methylation detection of tumor related genes,etc. In the high throughput sequencing technology, database and computer sequencing experiments are the key factor to decide the quality of DNA data. Based on IUumina sequencing instruments, the sample preparation method and sequencing running for PCR products were optimized,the results make us clearly understand the primary features of high,throughput sequencing techniques, and provide us important references of sequencing methods for low diversity DNA samples to address biological questions of interest.