Method and system for detecting oligonucleotide synthesis quality

Abstract

The invention belongs to the technical field of genes, and more specifically relates to a method for detecting the synthesis quality of oligonucleotide, which comprises the following steps: 1) synthesizing oligonucleotide, retaining an uncoupled 5 '-hydroxyl group in the synthesis process without closing, the oligonucleotide sequence comprising a sequence to be detected; 2) sequencing the oligonucleotide sequence synthesized in the step 1) to obtain sequencing data; and 3) performing gene mutation information analysis and calculation on the sequencing data obtained in the step 2) to obtain the proportions of different sequences in the oligonucleotide sequence in all the sequences and the proportions of correctness, deletion, mutation and insertion of basic groups at different positions. The method provided by the invention can accurately detect the correctness of the oligonucleotide base sequence, the synthesis efficiency of each step and the type and position of each wrong base.

Description

technical field [0001] The invention belongs to the field of gene technology, and more specifically, the invention relates to a method for detecting the quality of oligonucleotide synthesis. Background technique [0002] Oligonucleotides are usually composed of dozens of nucleotides and are a type of single-stranded nucleic acid molecule with a short sequence. Oligonucleotides can be used as primers, gene probes, synthetic gene base fragments, gene therapy-related drugs, etc., and are widely used in modern molecular biology research. A common method for the synthesis of oligonucleotides is solid-phase phosphoramidite chemical synthesis, which is still used by most commercial DNA synthesis companies. The synthesis method is a cyclic process of extending the nucleotide chain from the 3' end to the 5' end, and the nucleotide chain will be extended from the first protected nucleotide molecule immobilized on the surface. Among them, the immobilized carrier is mainly controlled ...

AI Technical Summary

Problems solved by technology

These methods are detected by the physical and chemical differences caused by the difference in the number of oligonucleotide bases, so the correctness of the oligonucleotide base sequence cannot be detected, and the correct sequence oligonucleotide and the wrong sequence oligonucleotide cannot be distinguished.

Secondly, in the process of detecting the synthesis efficiency, the above method can only detect the purity of the final product. Assuming that the synthesis efficiency of each step is the same, the synthesis efficiency of each step is reversed, and the average value of the synthesis efficiency of each step is obtained. Therefore, the method The accuracy is poor and cannot reflect the reaction efficiency of each step in the real synthesis process

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