Methodology
DNA immunization of mice by a gene gun- Detection of immune reactions with ELISA and ELISPOT assays and intracellular staining of cytokines
- Isolation of cells from tumors and their characterization with flow cytometry
- Analysis of gene expression – RT-qPCR, immunobloting, immunofluorescent staining
- Gene cloning and modification by mutagenesis
- Inactivation of cellular genes with the CRISPR-Cas9 system
- Next generation sequencing (scRNA-seq, WES)
Laboratory Equipment
- Gene gun (Bio-Rad) for biolistic immunization of mice with DNA vaccines
- AutoMACS (Miltenyi Biotec) for magnetic separation of cells
- GentleMACS octo (Miltenyi Biotec) for tissue homogenization and preparation of single cells
- Necessary equipment for cell cultures – laminar flow boxes, CO2 incubators, centrifuges, microscopes
- Necessary equipment for electrophoretic separation of DNA, RNA, and proteins – electrophoresis units, power supplies, blotting apparatus