Nucleic acid sequence sequencing linker and method of using same to construct sequencing library

Abstract

The invention provides a nucleic acid sequence sequencing linker and a method of using the same to construct a sequencing library. The sequencing linker includes a long Y primer and a short Y linker; the short Y linker and the long Y primer are connected to the two ends of an inserted fragment under sequencing respectively; the short Y linker has an upstream primer F and a downstream primer R which are partially complementary to each other; the long Y primer has an upstream primer P5 and a downstream primer P7; the short linker herein is used to perform tag identifying on different molecules in a same library, while the long Y primer is used to perform tag identification on different libraries. The nucleic acid sequence sequencing linker and the method herein are suitable for more accurately recognizing information of mutation sites, positive predicting capacity of a detection system can be improved greatly, and the problems in liquid biopsy application are solved.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a nucleic acid sequence sequencing connector and a method for constructing a sequencing library. Background technique [0002] Liquid biopsy makes up for the lack of tissue detection. Different patients with the same cancer type, different treatment stages of the same tumor patient, and different regions of the same tumor tissue have certain differences in the biological characteristics of the tumor. Liquid biopsy can not only overcome the heterogeneity of tissue detection, but also has the characteristics of simplicity, safety, non-invasive, real-time, etc., especially for patients who cannot undergo surgery or puncture, or where the tumor location makes sampling difficult. Methods that can make up for the limitations of tissue detection have played an important role in the real-time assessment of tumor targeted therapy and drug resistance monitoring in recent years. ...

AI Technical Summary

Problems solved by technology

However, due to the complex experimental process technology of NGS, some amplification and sequencing errors will inevitably be introduced in the process of library construction, target region capture, and sequencing. These errors are called background noise, and ctDNA detection often has a relatively high mutation frequency. Low, subject to background noise interference, low-frequency mutations from ctDNA samples are often submerged in background noise, resulting in false negative or false positive results, which limits the sensitivity and specificity of ctDNA detection

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