Method and system for determining individual gene mutation
An individual and genetic technology, applied in the field of determining individual gene mutations, can solve the problems of repeated sample consumption, large demand, and limited guiding significance, and achieve reduced sample volume and sequencing costs, reliable tumor genetic information, accurate and fast typing Effect
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[0292] (1) Tumor patient T30, diagnosed as hepatic differentiated cholangiocarcinoma, DNA and RNA were extracted from paraffin section of tumor tissue and frozen blood (normal tissue) of tumor patient T30, and passed the test.
[0293] (2) Targeted gene sequencing by chip capture: A total of 434 genes with high sensitivity to treatment were selected from the tumor gene database and its gene capture chip (AgilentSureSelect) was designed, and these 434 genes were performed on the sample DNA obtained in step (1). The exon part of the gene and the fusion part of 21 genes were captured, and the obtained DNA fragment was built and sequenced at a depth of 500x by the Illumina Hiseq sequencing system. The sequencing volume of a single sample was about 4.4G.
[0294] a. 3μg DNA sample was broken into 200-300bp fragments and purified.
[0295] b. End-repair and end-adding "A" were performed on the fragmented samples and purified.
[0296] c. Ligate the product obtained in step b with t...
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