Efficient RNA reverse transcription method and RNA and DNA synchronous library building method

Abstract

The invention relates to an efficient RNA reverse transcription method and an RNA and DNA synchronous library building method. DNA and RNA are mixed and then subjected to reverse transcription together, the RNA reverse transcription efficiency is improved, all reverse transcription products are directly and synchronously built into a library, an rhPCR amplification method is adopted in the librarybuilding process, the possibility of dimer generation is reduced, the risk of secondary library building is effectively avoided, and the operation is simpler.

Description

technical field [0001] The invention relates to the technical field of gene sequencing, in particular to a high-efficiency RNA reverse transcription method and a simultaneous RNA and DNA library construction method. Background technique [0002] Next-generation sequencing (NGS) technologies have revolutionized the field of genomics over the past decade. Each NGS run typically generates gigabytes of sequence information on hundreds of thousands to billions of DNA templates in parallel for a single sequencing run. The current cost for sequencing the human genome has reached the $1,000 benchmark. The low cost and high throughput of NGS technology have enabled the use of nucleic acid sequencing as a clinical tool. [0003] The genetic traits of organisms are mainly determined by nucleic acids, which include DNA and RNA. In recent years, with the development of molecular biology and various omics technologies, the comparative study of nucleic acid sequences has gradually becom...

AI Technical Summary

Problems solved by technology

In the traditional NGS library construction of DNA or RNA, DNA and RNA are constructed separately. This method has a single detection item, and the cost of consumables, labor and reagents is high

The current main method for detecting SNV sites and Fusion sites is to build separate libraries, because SNV detection is based on DNA, while Fusion detection is based on RNA. The former is to directly construct a DNA library, while the latter needs to be reverse-transcribed into cDNA first. Then construct the cDNA library, the two need to be operated separately, and then sequenced on the machine at the same time, the operation is cumbersome

Due to the need to construct two kinds of libraries, the library construction process is complicated, and the risk of library construction failure is relatively increased

At the same time, the library construction technology of amplicon sequencing usually uses a large number of primers, which can easily generate a large number of primer dimers, which will also increase the complexity of the operation and the demand for testing samples.

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