Plasmid DNA Isolation and Characterization Studies

Abstract

Objective: The use of gene therapy for targeted therapy in various genetic diseases has become more and more popular day by day. Various carrier vectors are used in gene therapy. The plasmid we are interested in in this study is used as a gene delivery vector. In this study, it was aimed to perform the isolation and characterization studies of the peGFP-N1 plasmid. In addition, necessary properties were investigated to make it a suitable gene carrier vector. Method: In this study, isolation was performed using the plasmid peGFP-N1 via classical method. Escherichia coli was used as expression medium in the isolation process. In order to calculate the amount of plasmid DNA in the studies, the spectral curve value of the plasmid DNA was calculated. Spectrophotometric methods were used to determine the amount and impurity of the isolated bacterial DNA. Results: In plasmid DNA isolation studies, it was observed that the yield was quite high and the contamination from protein or RNA was very low. In fact, the ratio of absorbance values (A1/A2) is very close to 1.8 as a result of the isolation III study shows that there is no contamination. Conclusions: The isolation and spectrophotometric study results of the peGFP-N1 plasmid showed that this plasmid is suitable and usable in terms of impurity and contamination risk. By making use of the results of this study, it will support the studies in which this plasmid can be used in an appropriate gene carrier system.