Research Specifications

The generation of bacterial competent cells is a key step in the cloning and genetic engineering process, which allows the introduction of foreign DNA into the cell. Classical methods such as calcium chloride (CaCl₂) treatment and electroporation have been used as standard approaches for competentization for many years. However, these methods face challenges such as low efficiency, sensitivity to environmental conditions, and limitations in the transfer of large-sized DNA. In recent years, novel technologies such as the CRISPR-Cas genome editing system and direct DNA injection (microinjection) have attracted the attention of researchers. These techniques can improve the efficiency and accuracy of genetic transformation by modifying the DNA uptake pathways or its physical transfer. In this article, a comparative study between classical and novel methods for the generation of bacterial competent cells is presented, and the advantages, disadvantages, and future prospects of each are summarized.