Method for sequencing high-flux circular RNA (ribonucleic acid)

Abstract

The invention discloses a method for sequencing high-flux circular RNA (ribonucleic acid), belonging to the field of molecular biology. The method comprises the following steps: artificially synthesizing exogenous linear ERCC-0004-RNA and exogenous circular ERCC-0013-RNA; performing quality control on the exogenous circular ERCC-0013-RNA; mixing the exogenous linear ERCC-0004-RNA and the exogenous circular ERCC-0013-RNA at an equal mole ratio to obtain a mixed exogenous RNA; adding the mixed exogenous RNA into total RNA of a to-be-sequenced sample to obtain a first mixture; removing ribosome RNA in the first mixture to obtain a second mixture; performing 3' tail end biotin marking on the second mixture, and removing lariat RNA and linear RNA marked with biotin to obtain a third mixture; removing linear RNA in the third mixture to obtain a fourth mixture; and performing standard high-flux transcriptome sequencing on the fourth mixture and performing biological analysis on sequencing data. By adopting the method disclosed by the invention, the expression of genes in an organism under a specific condition can be accurately reflected, and the cost is reduced.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method for high-throughput circular RNA sequencing. Background technique [0002] Most of the RNA in organisms is linear, but a small number of RNA end-to-end links form a circle, which is called circular RNA. As early as 1980, circular RNA was discovered for the first time, but for 30 years, due to the limitation of research methods, people knew little about circular RNA. Until recent years, with the rapid development of high-throughput sequencing technology, and Combined with the method of bioinformatics, large-scale transcriptome data was deeply mined, and researchers discovered a large amount of circular RNA information, which made circular RNA quickly become a new hotspot in the RNA world research, and then derived targeted It also greatly speeds up the discovery of circular RNAs and the analysis of their functions. In existing high-throughput transcriptome sequencing, b...

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Problems solved by technology

[0004] In order to solve the problem that the circular RNA in the total RNA cannot be accurately sequenced in the prior art, the embodiment of the present invention provides a high-throughput circular RNA sequencing method

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