对多个混合DNA或RNA序列进行基因测序的方法及装置

本发明公开了一种对多个混合的DNA或RNA序列进行基因测序的方法及装置，主要是在建库前给每个DNA或RNA模版序列两端加入随机标签。方法包括：将含有随机标签和接头的第一上游引物与第一下游引物和DNA序列混合，进行两个PCR循环的多重PCR扩增，纯化该PCR产物，获得第一DNA产物；将第一DNA产物与含有样品索引序列和接头的第二上游引物和第二下游引物混合，进行PCR反应，获得第二PCR产物；纯化后获得第二DNA产物；对第二DNA产物进行测序，获得每个DNA序列的测序结果。本发明通过引入随机标签到每个DNA分子，能够提高测序的准确性，显著减少测序的错误率，且准确测定每种DNA的拷贝数。

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.