Method for constructing transcriptome library

A construction method and transcriptome technology, applied in chemical libraries, biochemical equipment and methods, combinatorial chemistry, etc., can solve the problems of low comparison rate of sequencing data, avoid waste of human and financial resources, reduce sample volume, and simple operation Effect

Inactive Publication Date: 2015-05-20
BEIJING NOVOGENE TECH CO LTD
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[0005] The main purpose of the present invention is to provide a method for constructing a transcriptome library, so as to solve t

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  • Method for constructing transcriptome library
  • Method for constructing transcriptome library
  • Method for constructing transcriptome library

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[0023] 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 with reference to the accompanying drawings and examples.

[0024] As mentioned in the background technology section, the alignment rate of the sequencing data obtained after sequencing the transcriptome library constructed in the prior art is low. In order to improve this situation, in a typical embodiment of the present invention, the A method for constructing a transcriptome library, such as figure 1 As shown, the method comprises the following steps: S1, identifying whether there is contamination in the total RNA in the bacterial strain to be tested; S2, removing rRNA from the total RNA of the bacterial strain to be tested without contamination, to obtain mRNA and non-coding RNA (non-coding RNA) -coding RNA); S3, construct the transcriptome library...

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Abstract

The invention discloses a method for constructing a transcriptome library. The construction method comprises the following steps: S1, verifying whether total RNA in a strain to be measured is polluted; S2, removing rRNA from the total RNA of the unpolluted strain to be measured to obtain mRNA and non-coding RNA; and S3, using the mRNA and the non-coding RNA to construct the transcriptome library to obtain the transcriptome library. In the method, a step of controlling the quality of the total RNA is added to ensure that the RNA sample is not polluted, and a subsequent transcriptome library construction step is carried out to avoid the waste of manpower and financial resources caused by the polluted RNA and perfect the problem of low comparison rate of subsequent sequencing data. Few samples are needed in the verification step, the operation is simple, a result is obtained quickly and accurately, and the verification step has high throughput capacity, so that the verification step can be widely used in the transcriptome library construction step to verify the total RNA to the strain to be measured.

Description

technical field [0001] The present invention relates to the field of library construction, in particular to a method for constructing a transcriptome library. Background technique [0002] Using the second-generation high-throughput sequencing technology to sequence the transcripts (mRNA and non-coding RNA) of the biological sample to be tested can comprehensively and quickly obtain the information of all transcripts in a specific state of the biological sample to be tested. The molecular regulatory mechanism of the formation of different phenotypes of the biological samples to be tested can be revealed through transcriptome sequencing research. [0003] When the library is constructed by transcription, rRNA in the total RNA is usually removed first (it can be removed using the Ribo-zero kit), and then the RNA library construction kit in the prior art is used to construct the transcriptome library (such as using NEB Ultra Directional RNA Library Prep Kit for library constr...

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68C40B50/06
Inventor 曹志生李宗文王大伟李富威李明洲刘运超朱海浩
Owner BEIJING NOVOGENE TECH CO LTD
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