Gene panel for predicting new antigen load and detecting genome mutation
A genome and gene technology, applied in DNA mutation detection technology, bioinformatics, and biotechnology fields, can solve the problem of inability to meet the needs of multi-target high-throughput detection, lack of rapid, efficient and accurate prediction methods for neoantigen load, etc. question
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[0026] 1. Library preparation for next-generation sequencing
[0027] (1) Genomic DNA was extracted from samples (such as cancer tissue, blood or other types of cancer biopsies) using Qiagen DNeasy kit or QIAamp cell-free nucleic acid extraction kit (Qiagen);
[0028] (2) Use Qubit 3.0 Fluorescence Quantitative Instrument (Thermofisher) and Tapestation System (Agilent) to check the quantity and quality of extracted genomic DNA;
[0029] (3) Input 10-100ng of genomic DNA and use the NeoCura NGS library preparation kit for NGS library preparation;
[0030] (4) Use Qubit 3.0 Fluorometer and Tapestation system to check the quantity and quality of NGS library (see figure 2 ).
[0031] 2. Use the gene panel to capture the target DNA
[0032] (1) Use 100-500ng library for target capture;
[0033] (2) Hybridize with the gene panel probe at 65°C for 4-16 hours;
[0034] (3) Using Dynabeads TM M-270 streptavidin magnetic beads separate and purify the captured target DNA;
[003...
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