Abstract: It is important to use a simple protocol for Bacterial Ghosts (BGs) preparation. Recently, a new protocol has been introduced based on simple chemical compounds to prepare the BGs. The protocol has been given the name “Sponge-like” (SL). The used microbial strain was E. coli BL21 (CDE3) pLysS (Promega). Experimental Design (ED) has been used to optimize the preparation conditions. In general, to map the best conditions for BGs preparation, several steps have been followed and lead to a long protocol. There is a need for reducing the protocol steps. In this study and based on that strains within the same species are similar and aiming to introduce a simple BGs protocol, another E. coli recombinant strain, the E. coli JM109 (Promega) has been used. The optimum conditions obtained from the SL protocol were used. However, the MIC and the Minimum Growth Concentration (MGC) were determined. Light and electron microscope as well as the protein and the DNA contents were used to evaluate the BGs quality (BGQ). This study establishes a simpler protocol given the name “Sponge Like Reduced Protocol “SLRP”. The protocol proves to be suitable for BGs preparation from strains within the same species and could be used for native or recombinant antigens preparation.