Connected sequencing method for DNA by coupling and coding two rounds of signals

Abstract

The invention discloses a connected sequencing method for DNA by coupling and coding two rounds of signals. According to the method, the coding mode of markers is optimized so as to establish coupling correlation between two rounds of signals by virtue of a simple signal coding relationship without increasing the number of the markers, and the information of more than one base is detected in one round of sequencing reaction, thus forming a rapid, accurate, low-cost and high-throughput sequencing method for DNA. By coupling the two rounds of sequencing signals, the information of three bases can be detected, and the sequencing speed can be increased greatly, and under the condition that the connection efficiency is unchanged, the sequencing read length can be increased by 50%, and the sequencing cost can be reduced to a certain extent. In addition, because the two rounds of sequencing signals can be checked mutually, the accuracy of the sequencing result can be improved favorably.

Description

technical field [0001] The invention relates to a DNA link sequencing method using signal coupling coding, which is a method for realizing DNA sequence analysis, and belongs to the field of biotechnology. technical background [0002] In recent years, with the deepening of human understanding of genes, especially the 1000 Genomes Project and the continuous sequencing of various biological genomes, great changes have occurred in the research and application of biology and medicine. More and more research and detection methods are carried out at the level of genes, including studies on the differences between living organisms, the occurrence and development of diseases, and the interaction between drugs and living organisms. And all of this depends on the development and progress of genetic testing methods. How to quickly and efficiently detect and screen the differences between genes and between genomes and genomes has become one of the research hotspots in the post-genomic e...

AI Technical Summary

Problems solved by technology

These commercial sequencing systems have greatly improved the efficiency of DNA sequencing and reduced the cost of DNA sequencing. However, they still cannot meet the needs of life science research and medical testing in terms of sequencing cost, throughput, speed and accuracy.

The main reason is that the signal labeling method for detecting nucleic acid information is single and inefficient, or the labeling scheme is cumbersome and affects other technical parameters

For example, in DNA linkage sequencing methods, the general scheme is limited by the type of markers (such as fluorescent groups), and only one base can be determined for each ligation reaction. For example, two or more bases need to be determined in one ligation reaction. If there are more than two base information, the types of markers required will increase exponentially; in some schemes, researchers have developed a connection sequencing method based on signal combination coding, and two-dimensional or multi-dimensional Encoding, so as to realize the interpretation of two bases at a time without increasing the number of markers. However, this encoding method needs to interpret the presence or absence of 4-5 marker signals at most at one signal point, which requires the signal interpretation capability of sequencing equipment. Very high, greatly increasing the cost of sequencing equipment, and at the same time due to the interference between multiple markers, the accuracy of sequencing has been reduced to a certain extent

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