Molecular mass determination
The cell free supernatant of SB93 culture was treated with
butanol to extract bacteriocin (Ahmad, Iqbal, Haseeb, & Khan,
2014). Thin layer chromatography (TLC) of bacteriocin was performed in a solvent system of butanol, acetic acid and water in the
proportion of 4:1:1 (Ahmad et al., 2014).
Molecular mass of SB93 bacteriocin was determined by SDSPAGE with 5% stacking gel and 12% separating gel. After electrophoresis, one half of the gel was stained with Coomassie blue and
fixed in water containing 20% methanol and 10% glacial acetic acid
(Lu et al., 2014).
The other half was washed thrice with sterile water, overlaid
with V. cholerae culture and incubated at 37 C for 24 h.