RNA analyzing method of paraffine section tissue

Abstract

The invention provides an RNA analyzing method of paraffine section tissue. The analyzing method comprises the following steps of performing DNA degradation on the paraffine section tissue, and extracting RNA of samples; preparing a paraffine section sample nucleic acid library, and sequencing sample RNA; performing quality control on the sample data obtained by sequencing; comparing the sample data after quality control with a reference genome, and performing quality control on comparing results; and performing transcriptome assembling and transcript quantifying on the sample data after quality control of comparing results, and performing quantification analysis, gene difference expression analysis and fusion gene analysis on gene expression. The invention provides an index and detectionmethod for completely assessing RNA quality of the paraffine section tissue, so that RNA of the paraffine section tissue can be comprehensively assessed, and the assessment results are accurate and effective. An effective reference basis is provided for subsequent analysis accuracy.

Description

technical field [0001] The invention belongs to the field of second-generation high-throughput sequencing analysis, and in particular relates to a method for analyzing tissue RNA of paraffin sections. Background technique [0002] RNA sequencing (RNA-seq) is a sensitive and accurate method for quantifying gene expression. Next-generation high-throughput sequencing (NGS) has ushered in a new era for RNA-seq transcriptome analysis. The design of the broad-spectrum application process of RNA-seq involves sequencing technology, sample type, genome demand analysis, and computing resources. The analysis process is evaluated in terms of accuracy, computational speed, and the cost of analysis. [0003] Gene expression profiling of tumor samples is a powerful biomarker for identifying prognosis and prediction. To date, transcriptional profiling has been performed on a large number of cancer frozen tissue samples. However, formalin-fixed paraffin-embedded tissue (FFPE) is a more w...

AI Technical Summary

Problems solved by technology

FFPE samples generally have severe degradation, chemical modification, cross-linking of nucleic acids and proteins, and variability in tissue processing and processing. These molecular changes will directly affect data quality and bring several major problems, such as sample degradation leading to sequencing data comparison quality Low, resulting in more soft-cut sequences, more repetitive sequences, and formaldehyde fixation leading to random C-T conversion of nucleic acids, which make FFPE-separated nucleic acids incompatible with downstream high-throughput molecular techniques

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