高通量测序DNA文库定量质控技术研究

DNA文库定量的准确与否直接影响测序数据质量的好坏,因此,开展文库定量质控技术研究非常重要。针对Illumina高通量测序平台的DNA文库定量,建立了基于SYBR Green荧光染料嵌合的高特异性数字PCR绝对定量方法,用于DNA文库定量标准物质的定值;并对制备的文库定量标准物质的适用性进行了验证。结果表明制备的5个水平的文库定量标准物质线性良好,可以作为Illumina平台DNA文库定量的标准曲线。

Research on Characterization Method for Library DNA Reference Material for Next Generation Sequencing

Next generation sequencing (NGS) have been increasingly used in many applications. The accuracy of quantification of DNA library can directly affect the quality of the sequencing data. Thus, it is very important to develop DNA library quantification quality control method. A SYBR Green dye based digital PCR method was established to characterize the DNA library quantification reference material for Illumine NGS platform. The suitability of the proposed reference material was evaluated. The result showed that it is fit for the purpose of usage as standard curve to quantify the DNA library for illumina NGS.The results showed that the five levels of library quantitative reference materials have good linearity, and can be used as the standard curve to quantify the DNA library for Illumina NGS.