TRANSFORMATION

STEPS MATERIALS N AM E CATAL OG # VEN DOR LB Broth J106-2KG Transformation V.4 Rengin Yamur Akbiyik , Sevval Uysalcan Istanbul Bilgi University Works for me dx.doi.org/10.17504/protocols.io.2nwgdfe Rengin Yamur Akbiyik Istanbul Bilgi University May 07, 2019 1 1 1 Transformation is a direct modification of the genotype of a cell from a different one by extracellular applications using recombinant DNA techniques. Transformation refers generally to the integration of exogenous DNA into the cell and its integration into the genome. Before starting the transformation process, the gene of interest is ligated with the plasmid vector via the ligase enzyme. This process is called ligation. Generally, nucleic acids or plasmids cannot enter into bacterial cells by themselves. The stimulatory effect is required for that purpose. This means that the cell membranes to be transformed must be pre-arranged. Bacteria that can contain free DNA are called competent bacteria. Some bacteria are highly competitive in normal growth conditions; but some must be treated with chemical or physical methods to gain competitive properties. Competent cells which are cells are generally stimulated with calcium chloride about chemically and the cell membranes are arranged such that the plasmid vector containing the gene of interest can harbor the vector. Some bacterial cells are lasting competent, however; most of them need to be influenced for being competent. There are two ways about this transformation process that; • Chemical Transformation • Electrical Transformation (Electroporation) In general, chemical transformation is used. In chemical way, some special chemicals are used to open the pores which are found in cell membrane. This is because, opening the pores represents the availability of being permeable. Most of the time, divalent ions like Ca++ are used with assistance of heat-shock.