In Silico Analysis and Validation of ymcA Gene of Bacillus Subtilis

Abstract

This study aimed to analyze and validate the ymcA gene of Bacillus subtilis in silico. The bacterial strains used were B. subtilis 2049 (T) and B. subtilis HS, obtained from the National Center for Microbial Resource, Pune. The media used were LB medium and Mineral Salt Medium (MSM). The biofilm formation assay was performed in a microtiter plate, with each well inoculated with 980 µl of MSM and 20 µl of test cultures. The plate was incubated for 120 hrs at 37°C following phosphate buffer assay. The contents of the plate were removed by gently tapping at the bottom of the plate, rinsed with phosphate buffer saline, and de-stained using 70% ethanol for 15 min. The genomic DNA of B. subtilis 2049 (T) was extracted using the Chloroform: Isoamylalcohol method. The supernatant was taken in a sterile tube and stored at -80°C until further use.