Gene set for liver cancer detection and detection and design method of panel of gene set
A liver cancer and gene technology is applied to the gene set for liver cancer detection and its panel detection design. The application of the gene set in the gene chip can solve the cumbersome process of information analysis method, the omission of important sites, and the single technical process, etc. problems, to achieve the effect of strong design controllability, low detection and analysis costs, and high overall coverage
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preparation example Construction
[0052] As a preferred embodiment, the preparation method of the gene panel of the present application mainly includes the following steps:
[0053] 1. Screening of liver cancer information and related genes. Including the collection of at least three aspects of liver cancer information and its related genes, the first aspect, the collection of liver cancer information and its related genes obtained from the sequencing of liver cancer patient samples in the local database; the second aspect, the collection of liver cancer information and its related genes disclosed in the public database. Genes; the third aspect is to collect liver cancer information and related genes disclosed in the literature; merge and remove redundancy from the liver cancer information and related genes collected in the three aspects, and determine the standard gene name through HGNCapproved Official Symbol to form a gene set ;
[0054] 2. Detection target area selection. Based on the gene set obtained i...
Embodiment 1
[0069] Example 1 Panel sequencing
[0070] 1) Sample collection
[0071] According to the collection standards of the "Chinese Standards for the Diagnosis and Treatment of Primary Liver Cancer", medical institutions collected samples from 110 liver cancer patients.
[0072] 2) Sample DNA extraction, library preparation
[0073] For individualized gene detection of liver cancer, liver cancer tumor FFPE is used as the experimental group for each sample, and the blood samples collected at the same time are used as the control group. Considering the influence of label joints, hybridization, and data volume utilization, the number of databases established is evaluated, and the number of databases established once is confirmed. 4 pairs of library samples are the best. Use FFPE Nucleic Acid Extraction Kit and Blood Cell Extraction Kit for sample extraction respectively, and use Qubit DNA Fluorescence Quantitator and Qubit dsDNA HS Assay kit for quantitative detection. The extractio...
Embodiment 2
[0087] Example 2 Data analysis after getting off the machine
[0088] The information analysis process is used for the paired sample analysis of the liver cancer project, taking the off-machine Fastq file of the sequencing platform (such as BGI-Seq500 in this example) as input, and obtaining The final mutation result and the basic statistical information of the data are displayed.
[0089] This protocol was used for the paired-sample analysis of the liver cancer project. For the fastq after the BGI-Seq500 is off the machine, it is first filtered by SOAPnuke to filter out the reads with a proportion of low-quality bases (≦10) greater than 50%, or a proportion of N (undetermined bases) greater than 10%. , and count relevant indicators before and after filtering, including initial data volume, effective data volume, CG content, N content, Q20, Q30, etc.; compare the clean fastq obtained after filtering to the human reference genome hg19, and this step is realized by paligner; A...
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