μl of a loading buffer (0.4% Bromo-phenol Blue, 0.4% xylene
cyanole and 5 ml of glycerol) and was analyzed directly on 1.5%
agarose gels in 0.5X TBE buffer. Electrophoresis was done for
about 3 h at 60 volts. The GeneRulerTM 1kb ladder plus (Fermentas
Inc, Hanover, MD) was used as a molecular size standard to compare
the molecular weights of amplified products. Visualization of
the amplified bands was done by staining with ethidium bromide (10
mg/ml) for 20 min and de-staining with double distilled water for 20
min followed by transillumination under short wave UV light (Alpha
Innotech).