High-quality 3 'RNA-seq library building method and application thereof

Abstract

The invention develops an efficient 3 'RNA-seq method, namely a simplified poly (A) anchoring sequencing (SiPAS V2). According to the method, a next-generation sequencing linker in the library is specifically turned, so that a non-poly (T) end of the library is read at an R1 end during sequencing, and the method is more suitable for a standard PE150 sequencing format. By evaluating the comprehensive performance of the SiPAS V2 in hexaploid wheat, it is proved that the SiPAS V2 has high sensitivity, accuracy and repeatability when gene expression is quantified.

Description

technical field [0001] The invention relates to a high-quality 3'RNA-seq library building method and use thereof, which can be used for accurate detection of gene expression levels in large-scale high-throughput populations. Background technique [0002] RNA sequencing (RNA-seq) is a key technology in modern biological research, which transforms the study of many species from a single genome level to a multidimensional omics level, thereby effectively improving our understanding of biological genomics. In the past few years, whole-genome sequencing of many crops has been completed, generating a large amount of genome data, for example, the genetic variation map of wheat, maize, rice, cassava, potato and soybean, etc. At the same time, many important crops have also assembled High-quality pan-genome. The study of large amounts of genomics data has created a research vacuum that needs to be populated with large amounts of transcriptome data to help decode genome function. Th...

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