RNA analyzing method of paraffine section tissue

A technology of paraffin sectioning and analysis methods, applied in sequence analysis, biochemical equipment and methods, instruments, etc., can solve the problems of low quality of sequencing data comparison and technical incompatibility, and achieve accurate and effective evaluation results

Pending Publication Date: 2020-01-14
深圳吉因加医学检验实验室
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  • Abstract
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  • Application Information

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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 leadi

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  • RNA analyzing method of paraffine section tissue
  • RNA analyzing method of paraffine section tissue
  • RNA analyzing method of paraffine section tissue

Examples

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Embodiment 1

[0041] like figure 1 As shown, a method for RNA analysis of paraffin section tissue includes the following steps: DNA degradation is performed on paraffin section tissue, and sample RNA is extracted; a nucleic acid library of paraffin section samples is prepared, and based on the library, the sample RNA is sequenced; The sample data obtained by sequencing is subjected to quality control, and rRNA data is removed; the sample data after the quality control is compared with the reference genome, and the comparison results are subjected to quality control; the sample data after the quality control is subjected to quality control; Transcriptome assembly and transcript quantification are used to quantitatively analyze gene expression; based on the transcript quantification results, gene differential expression analysis is performed; at the same time, fusion gene analysis can also be performed using the method described in this example. The software for each step and its result files...

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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...

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Application Information

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IPC IPC(8): C12Q1/6869C12Q1/6886G16B30/00
CPCC12Q1/6869C12Q1/6886G16B30/00C12Q2600/158C12Q2535/122C12Q2525/191C12Q2531/113
Inventor 黄毅易鑫吴玲清刘久成王长希李俊
Owner 深圳吉因加医学检验实验室
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