Transcriptome sequencing method

Abstract

The invention discloses a transcriptome sequencing method, which comprises the following steps: (1), preparing total RNA, enriching mRNA, randomly interrupting RNA, synthesizing cDNA fragments from RNA fragments by using random primers and reverse transcriptase, repairing tail ends of the cDNA fragments, connecting the cDNA fragments to a sequencing joint, preparing a sequencing library and carrying out sequencing; (2) preparing total RNA, removing rRNA, enriching mRNA, randomly interrupting RNA, synthesizing cDNA fragments from RNA fragments by using random primers and reverse transcriptase,repairing tail ends of cDNA fragments, connecting the cDNA fragments to a sequencing joint, and preparing a sequencing library and carrying out sequencing; and (3) preparing total RNA, then connectingSmall RNA, 3' end and 5' end joints, carrying out reverse transcription synthesis to obtain cDNA fragments, and preparing a sequencing library and carrying out sequencing. According to the transcriptome sequencing method provided by the invention, clinical samples are used for positioning and quantitative expression verification of transcriptomes, finding evidences of the transcriptomes and clinical relevance, and evaluating clinical values, so that new clues are provided for the pathogenesis of liver cancers and the research of liver cancer mechanisms.

Description

technical field [0001] The invention relates to the fields of gene transcriptomics and proteomics, in particular to transcriptome sequencing. Background technique [0002] One of the most important fields in biological and medical research is the observation of the structure, function and regulation of biological systems and life processes. However, in the past few hundred years, biologists have been focusing on the expression changes and functions of individual genes or proteins in biological systems, but cannot study the changes in living systems from a global and holistic perspective. With the advancement of medicine, people have found that many diseases, especially cancer, are often caused by the synergistic effects of multiple factors, multiple genes, and multiple pathways. This requires a technology and means that can comprehensively, dynamically and systematically study the life system, so the concept of "omics" came into being [3] . However, with the completion of...

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Problems solved by technology

However, with the completion of the Human Genome Project, it was found that only from the perspective of genomics can not completely correctly predict the splicing, splicing, and the start and end positions of open reading frame codons during translation and post-translation various modifications of

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