The present research study have been focused on the analysis of plasmid pattern and restriction digestion in Methicillin Resistant Staphylococcus aureus (MRSA) and Coagulase Negative Staphylococci (CoNS) in clinical isolates. A total of 72 Sputum Samples were collected from the hospitals in Namakkal and Salem District, Tamil Nadu, India. The collected samples were plated on different media, namely Nutrient agar medium, Mannitol salt agar medium, Blood agar medium, MacConkey agar medium, DNAse Agar medium and Hichrome MeReSa agar medium and incubated at 37ºC for 12-24 hours. The incubated agar media plates were studied for Morphological characteristics, Staining reaction and Biochemical characteristics. The Plasmid DNA was obtained from the isolated Staphylococcal species and the restriction digestion was carried through EcoRI and Hind III restriction enzymes.