What is the transfection reagent?
Transfection Reagent Basics Transfection is the process of introducing nucleic acids into eukaryotic cells by nonviral methods. Using various chemical or physical methods, this gene transfer method enables the study of gene function and protein expression in a cellular environment.
What is cacl2 transfection?
The calcium phosphate transfection method for introducing DNA into mammalian cells is based on forming a calcium phosphate-DNA precipitate. The DNA is mixed directly with a concentrated solution of CaCl2, which is then added dropwise to a phosphate buffer to form a fine precipitate.
What is calcium phosphate transfection?
Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. A DNA−calcium phosphate co-precipitate forms, which adheres to the cell surface and is taken up by the cell, presumably by endocytosis. Glycerol shock may increase the uptake of DNA in some cell types.
What is the function of transfection reagent?
Upon cellular uptake, transfection complexes are sequestrated into intracellular vesicles. Our transfection reagents are able to induce the release of the nucleic acids into the cytoplasm through vesicle membrane rupture or fusion.
What is transfection procedure?
Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. Transduction is often used to describe virus-mediated gene transfer into eukaryotic cells. The word transfection is a portmanteau of trans- and infection.
What is transfection method?
Transfection is the process of introducing nucleic acids into eukaryotic cells by nonviral methods. Using various chemical or physical methods, this gene transfer technology enables the study of gene function and protein expression in a cellular environment.
What is DEAE dextran?
Diethylaminoethyl (DEAE)-dextran is a polycationic derivative of the carbohydrate polymer dextran, and it is one of the first chemical reagents used to transfer nucleic acids into cultured mammalian cells (Vaheri and Pagano, 1965).
What is calcium phosphate method?
Abstract. The calcium phosphate transfection is a widely used method for introducing foreign DNA plasmids into cells. Mechanisms underlying this transfection method are not yet defined; however, DNA-calcium phosphate precipitates are internalized by the cells and DNA is efficiently expressed in almost all cell types.
What is the goal of transfection?
The main purpose of transfection is to study the function of genes or gene products, by enhancing or inhibiting specific gene expression in cells, and to produce recombinant proteins in mammalian cells [3].
What are the two types of transfection?
Generally, the methods can be divided into two categories: non-viral and viral.
How do you confirm transfection?
Determining the number of positive cells within a transfected cell population can be done through microscopy and flow cytometry. Finally, confirming localization of your protein of interest can be done by microscopy.