The ultrapure genomic DNA Spin Miniprep (Medox Biotech India) kit was used for fast isolation of genomic DNA. The quality of DNA thus obtained was measured using a nano-drop technique in the Department of Biomedicine of Sri Ramachandra University. The primers and probes specific for S mutans were manufactured by Bangalore Genei (Shushruti Nagar, Bangalore, India). The oligonucleotide primers used were Sm F5 50-AGC CAT GCG CAA TCA ACA GGT T and Sm R4 50CGC AAC GCG AAC ATC TTG ATC AG. The samples were then taken to the Central Research Foundation, Sri Ramachandra University, where the real-time polymerase chain reaction was done (model 7900HT; Applied Biosystems, Invitrogen BioServices India Pvt Ltd, Whitefield, Bangalore, India) using SYBR green assay for relative quantification of the bacteria in the samples.