Quality control for some frequently used cell lines: DNA sequence analysis of cytocrome B and cytochrome oxidase subunut I genes

In this thesis, HeLa, Cervix epithelloid carcinom (Human), HL-60, Human peripheral blood promyelocytic leukemia (Human), L929, Mouse C3 / An connective tissue (Mouse), MDCK, Madin Darby canine kidney (Dog), Neuro-2a, Mouse neuroblastoma (Mouse) cell lines were checked for contamination, their identity was verified and their clonality was determined. Theoretical approach and the outline of the method, in this study, DNA sequence analysis of mitochondrial cytochrome b (CYTB) and cytochrome c oxidase subunit I (COI) genes amplified from the DNA of cultured cells was aimed. These genes have stable sequences recognized by universal primers and a number of sequence variants to provide identification at the same time. In the thesis, the results of the comparison with the reference sequence in the database for the CYTB gene region were found to be 97% similar for "HL-60" and 98% for "HeLa, L929, MDCK, Neuro-2a" cell lines. For the COI gene region, these similarity rates were found to be 95%, 99%, 96%, 96% and 98%, respectively. In this context, DNA sequence analysis results obtained provided an acceptable indicator and confidence in the quality of "HeLa, HL-60, L929, MDCK, NA" cell lines used in many studies. At the same time, an evaluation was made in terms of mycoplasma contamination and no mycoplasma contamination was found. Targets and scientific, technological and socio-economic contributions: The determination of the characterization of cell lines used in experimental studies is important in terms of the reliability of the results of the research carried out with these cells. With the completed thesis, the identity of the cell lines that constitute the main source of the studies has been revealed, and this identification has revealed data that will contribute to the precision and reliability of the cell culture studies.