Molecular detection of coagulase (coa) gene

The primers used for amplification of coa gene of S. aureus are coa F: ATAGAGATGCTGGTACAGG and coa R: GCTTCCGATTGTTCGATGC [10]. The PCR amplification was performed in 25 µl in 0.2 ml thin-walled PCR tubes (Eppendorf, Germany). The PCR mixture contained a final concentration of 2.5 µl of 5X coloured buffer, 2.5 µl of 25 mM MgCl₂, 0.25 µl 25 µM concentration of each primer (Table-1), 0.5 mM concentrations of each 2’-deoxynucleoside 5’-triphosphate, and 2 U of Taq DNA polymerase (Promega, USA). The amplification cycles in Mastercycler Gradient Thermal cycler (Eppendorf, Germany) consisted of 94°C for 45 s, followed by 30 cycles of 94°C for 20 s, 57°C for 15 s, and 70°C for 15 s and final extension at 72°C for 2 min.

S. aureus isolates obtained from different samples using nuc gene PCR.

S. aureus=Staphylococcus aureus, PCR=Polymerase chain reaction