PLASMID CURING ASSAY ON E. coli
Plasmid Curing is the process where the plasmid of a bacterial sample is degraded with the help of chemical agents but before moving to curing, we need to have a basic idea of what plasmid is. We have heard of bacteria resistant to antibiotics. This happens due to the presence of plasmid / extra-chromosomal DNA which is found in the cytoplasm of the bacterial cell. This aids the bacteria to survive in a harsh environment. Bacterial plasmids contain genes which cater to antibiotic or heavy metal resistance. We can also insert genes in a plasmid to produce certain antibiotics or chemicals for other research purposes. Sometimes, certain bacterial species contain plasmids which can be very obscure and curing such plasmids is important to understand whether a particular trait is conferring from the genetics DNA or the plasmid DNA as the function of the plasmids remains elusive many times. So in order to understand such cases, plasmid curing is done with the help of various chemical agents like Acridine Orange, EtBr, etc. These act as an intercalating agent for DNA sequences that are used to degrade the plasmid DNA as plasmids contain mobile genetic elements which help the bacteria in their diversity and adaptability. So in this project, we did an assay on plasmid curing with the help of Acridine Orange and EtBr on Amoxicillin and Tetracycline resistant E. coli.