Method for preparing double-stranded RNA sequencing library

Abstract

The invention discloses a method for preparing a double-stranded RNA sequencing library, and an application of the method in biological genetic analysis. The method comprises the following steps: 1) randomly interrupting: carrying out random interruption treatment on to-be-detected double-stranded RNA to obtain a double-stranded RNA fragment; 2) optionally repairing an end: performing end repair on the double-stranded RNA fragment obtained in step 1) to obtain an end-repaired double-stranded RNA fragment; 3) performing linker connection: connecting the double-stranded RNA fragment in step 2) with a linker to obtain a double-stranded RNA fragment containing linkers at two ends; and 4) amplifying and purifying: by taking the double-stranded RNA fragment containing the linkers at the two endsin step 3) as a template, amplifying to obtain a double-stranded DNA product, and purifying to obtain the sequencing library. The library construction method is simple, convenient and rapid, and moreraw materials are saved.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a double-stranded RNA sequencing library. Background technique [0002] RNA virus is a kind of virus, such as hepatitis C virus, Japanese encephalitis virus, all influenza viruses, etc. Generally, the genetic material (ribonucleic acid) of RNA viruses is single-stranded (ssRNA, single-stranded RNA), and some genetic material is double-stranded (dsRNA, double-stranded RNA). Double-stranded RNA viruses are named because their nucleic acid is complementary double-stranded RNA. dsRNA viruses have two characteristics: one is that the viral genome is mostly double-stranded RNA molecules with 10-12 segments; the other is that the virus has a double-layer capsid and no envelope. The virus's double-stranded RNA transcribes mRNA under the action of the virus's own RNA polymerase, and then translates the early or late protein. When double-stranded RNA is replicated, it m...

AI Technical Summary

Problems solved by technology

[0003] At present, kits of many brands at home and abroad can realize single-stranded RNA library construction for high-throughput sequencing, but for some samples containing double-stranded RNA, there is no convenient and fast double-stranded RNA library construction for high-throughput sequencing on the market. Quantitative sequencing plan

Qi Zheng et al. (Genome-Wide Double-Stranded RNA Sequencing Reveals the Functional Significance of Base-Paired RNAs in Arabidopsis, 2010) disclosed a method for constructing a double-stranded RNA sequencing library, which fragments the double-stranded RNA and makes the RNA single chaining, and then sequentially connect sequencing adapters at both ends of the single-stranded RNA, and then obtain a DNA sequencing library through reverse transcription reaction. Many, it takes a long time to build a database

Images