Application in Next-Generation Sequencer (NGS) Library Quality Control (QC)

Introduction

Due to technological advancements in next-generation sequencers, they are being used for a wider range of applications, such as for de novo sequencing or for the analysis of mutations, exomes, and expressions. Furthermore, it is possible to select the model based on the capacity required for data analysis and data processing, which has increased their use rapidly. However, to obtain good sequencing results, regardless of the application or model, it is necessary to have a good understanding of the size distributions and concentrations in the NGS library. Therefore, quality control (QC) of that information is essential when using NGS systems. Previously, agarose electrophoresis was used to confirm size distributions in the NGS library, and a real-time PCR system or fluorescence spectrophotometer was required to confirm concentrations. However, the processes required, from library preparation to QC, involved a series of tedious manual operations. Therefore, there was a need for a quick, simple, and low-cost QC method. In particular, with the maturation of simultaneous multianalyte sequencing technology using index tag sequences, the number of libraries subject to quality control has been increasing, which has further increased the demand for such QC methods. This article describes an example of mouse RNA sequencing analysis that solves this problem by using an MCE-202 MultiNA automatic electrophoresis system for NGS library QC.

July 16, 2014 GMT