Method and device for gene sequencing of plurality of mixed DNA (Deoxyribonucleic Acid) or RNA (Ribonucleic Acid) sequences

Abstract

The invention discloses a method and a device for gene sequencing of a plurality of mixed DNA (Deoxyribonucleic Acid) or RNA (Ribonucleic Acid) sequences, which is mainly characterized by adding random labels at two ends of each DNA or RNA template sequence before establishing a bank. The method comprises the following steps of: mixing a first forward primer containing the random label and a connector, a first reverse primer and the DNA sequences, performing multiple PCR (Polymerase Chain Reaction) amplification on two PCR circulations, and purifying the PCR product to obtain a first DNA product; mixing the first DNA product, a second forward primer containing a sample indexing sequence and the connector, and a second reverse primer, performing PCR reaction to obtain a second PCR product; purifying to obtain a second DNA product; and sequencing the second DNA product to obtain a sequencing result of each DNA sequence. According to the method disclosed by the invention, by introducing the random labels to each DNA molecule, the sequencing precision is improved, the error rate of the sequencing is obviously reduced, and the copy number of each DNA is precisely detected.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a method and device for gene sequencing of multiple mixed DNA or RNA sequences. Background technique [0002] In the immune cell population, the diversity of T lymphocyte receptor molecules (TCR, T cell receptor) and B lymphocyte receptor molecules (BCR, B cell receptor) is hundreds of thousands to millions. The α and β chains that make up TCR or BCR not only have different structures, but the formation of each chain also undergoes the combination of many gene segments in different regions of V, J, or V, J, and D. [0003] When studying the diversity of TCR and BCR, it is necessary to sequence many gene fragments in the V, J or V, J, D regions of TCR and BCR. In general, existing sequencing technologies can be adapted to general gene sequencing requirements. However, for studying the diversity of TCR and BCR, the error rate of the existing sequencing technology is too hi...

AI Technical Summary

Problems solved by technology

However, for studying the diversity of TCR and BCR, the error rate of the existing sequencing technology is too high, so that it is impossible to distinguish the subtle differences among the numerous gene fragments based on the obtained sequencing results

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