A dna sequencing method comprising nucleotides and 3' end reversibly blocked nucleotides

Abstract

A DNA sequencing method that includes nucleotides and 3'-end reversibly blocked nucleotides. A single sequencing reaction is performed simultaneously by two different nucleotides, X and Y*, where X is an unblocked nucleotide at the 3'-end , Y* is a nucleotide that is reversibly blocked at the 3' end. According to the quantitative relationship between the number of detected molecules produced by two different nucleotides in the sequencing reaction and the number of synthesized nucleotides N, the sequencing information of a single sequencing reaction includes (N-1) specific base X and 1 or 0 Code XY. The whole sequencing includes performing at least two sets of sequencing reactions on the same template; finally, by comparing the two sets of sequencing information, the specific base sequence of the nucleic acid fragment to be tested is determined. Three sets of sequencing can also be performed, and the specific base sequence of the nucleic acid fragment to be tested can be determined by comparing the sequencing information obtained from the three sets of sequencing reactions, thereby further improving the accuracy of sequencing.

Description

technical field [0001] The invention belongs to the field of biotechnology, and is a method for realizing high-throughput determination of nucleic acid sequences, in particular to a DNA sequencing method comprising nucleotides and 3' end reversibly blocked nucleotides and its application. Background technique [0002] With the development and completion of the Human Genome Project and various model organism genome projects, human beings have entered the post-gene era, which has had a huge impact on contemporary biological research and medical research, and the related disciplines of molecular biology have been rapidly developed. develop. It will become possible to understand the differences of life, the law of disease occurrence and development, and the interaction between drugs and living organisms at the genetic level. Drastically reducing the cost of DNA sequencing will greatly promote research in life sciences and medicine, and even bring about revolutionary changes. A...

AI Technical Summary

Problems solved by technology

[0006] When only one nucleotide synthesis occurs in this sequencing reaction, it can be a synthesis reaction of X or a synthesis reaction of Y*, but it is unknown which nucleotide synthesis reaction occurs, so Only one code (XY) can be used to represent the sequencing information of this nucleotide;

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