Construction method of RNA methylation sequencing library

Abstract

The invention provides a construction method of an RNA methylation sequencing library. The construction method comprises the following steps: (S1) carrying out fragmentation on an RNA sample, so as to obtain fragmented RNA; (S2) carrying out co-immunoprecipitation on the fragmented RNA by virtue of an RNA methylation specific antibody, so as to obtain precipitates; (S3) digesting the precipitates by virtue of protease, so as to obtain a digestion product; (S4) extracting RNA from the digestion product, and carrying out reverse transcription on extracted RNA, so as to obtain cDNA; and (S5) constructing a sequencing library by virtue of the cDNA, so as to obtain the RNA methylation sequencing library. The digestion of protease is stronger than the competitive elution of analogues, so that the yield of RNA can be increased, the noise generated due to the introduction of methylation site analogues is avoided, the problem of high noise of constructed libraries in the prior art is solved, the library construction quality is improved, and therefore, the utilization rate of valid data output by the sequencing library is increased.

Description

technical field [0001] The present invention relates to the field of RNA sequencing, in particular to a method for constructing an RNA methylation sequencing library. Background technique [0002] Epigenetics refers to the heritable changes in gene expression and regulation without changing the genomic DNA sequence, and its phenomena mainly include DNA methylation, histone modification, genome imprinting and RNA modification. Among them, RNA methylation modification widely exists in eukaryotic organisms such as mammals and plants, and the main forms are m 7 G, N m 、m 5 C, m 6 A and m 6 A m . Different forms of RNA methylation play a vital role in the regulation of biological growth and development, such as m 7 G participates in translation modification, transfer, splicing, etc. of mRNA, N m Methylation is often used as a molecular marker for host cells to identify their own mRNA and foreign mRNA, and experiments such as gene knockout have shown that m 5 C, m 6 A pl...

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Problems solved by technology

[0021] The main purpose of the present invention is to provide a method for constructing an RNA methylation sequencing library to solve the problem of high noise in the library constructed in the prior art

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