0.75 ml of the above standard solution was diluted with 10.5 ml of distilled water to make a equivalent of 10 µg concentration. 150 µg and 10 µg discs were prepared by impregnating 20µl standard solutions. 6. Polymerase chain reaction (PCR) reagents Life technologies reaction (PCR) reagents 10 X PCR buffer (200 mM TrisHCL, pH 8.4, 500 mM KCL)
Control primer mix
50 mM MgCl2
Sterile water
10 mM dNTP mix
Silicone oil
Taq DNA polymerase
Molecular weight marker
Reaction mixture for aerA Reagents
Quantity
10 X PCR buffer
2.5 µl
50 mM MgCl2
1.5 µl
2.5 mM dNTP
1.5 µl
Primer 1 (aerA)(10 pm/µl)
1.0 µl
Primer 2 (aerA)(10 pm/µl)
1.0 µl
Taq polymerase
0.2 µl
Template DNA
2.0 µl
Distilled water
15.5 µl
ABBREVIATIONS A APW
Aeromonas Alkaline peptone water