Construction method of RNA methylation sequencing library

A construction method and technology for sequencing libraries, applied in chemical libraries, combinatorial chemistry, library creation, etc., can solve the problem of high library noise

Inactive Publication Date: 2016-05-25
BEIJING NOVOGENE TECH CO LTD
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[0021] The main purpose of the present invention is to provide a method for constructing an RNA methyl

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  • Construction method of RNA methylation sequencing library
  • Construction method of RNA methylation sequencing library
  • Construction method of RNA methylation sequencing library

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[0035] It should be noted that, in the case of no conflict, the embodiments in the present application and the features in the embodiments can be combined with each other. The present invention will be described in detail below in conjunction with examples.

[0036] As mentioned in the background technology section, the immunoprecipitation-based RNA methylation library construction method in the prior art has the defect of high noise resulting in relatively low library quality. In order to improve this defect, in a typical implementation of the present invention In the method, a method for constructing an RNA methylation sequencing library is provided, the construction method comprising: step S1, fragmenting an RNA sample to obtain fragmented RNA; step S2, using an RNA methylation-specific antibody to fragment Co-immunoprecipitating the RNA to obtain a precipitate; Step S3, using protease to digest the precipitate to obtain a digest; Step S4, extracting RNA from the digest and...

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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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Application Information

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IPC IPC(8): C40B50/06
CPCC40B50/06
Inventor 邓腊梅朱海浩李明洲宋彬彬齐鑫
Owner BEIJING NOVOGENE TECH CO LTD
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