The DNA damage activity of CFCS was determined onE. coli
pUC19 plasmid vector DNA. Plasmid DNA was oxidized with H2O2,
treated withUV radiation and checked on 1% agarose according to a
method described by Jakhar, Paul, Park, Han,&Kang (2014). In brief,
the experiment was performed in a 10 ml volume in a microfuge
tube containing 175 ng of pUC19 plasmid DNA in 1x TE buffer
(10 mM TriseHCl and 1 mM EDTA, pH 8.0). H2O2was added at a
final concentration of 33 mM with or without CFCS (3ml). The re-action was initiated by UV irradiation and continued for 35 s on the
surface of a UV trans-illuminator with an intensity of 22,500 mW/cm2
at 280 nm at room temperature. After irradiation, the reaction
mixture (10ml) along with gel loading dye (6x) was loaded on 1%
agarose gel for electrophoresis. Untreated pUC19 plasmid DNA was
used as a control in each run of gel electrophoresis. Gel was stained
with ethidium bromide and photographed on a Gel Doc system