First Step: PCR Amplification
Starting with DNA extracted from the environmental isolate, Polymerase Chain Reaction (PCR) is performed to amplify just a small region of the DNA used to identify the isolate. For bacteria, the 16S ribosomal RNA (rRNA) gene sequence is used. The 16S rRNA sequence is common across all bacterial species, and can be used to study the phylogenetic relationship among bacteria. Typically for MicroSEQ ID workloads, the first 500 nucleotide bases (the A, C, G, and T of DNA) of the sequence are sufficient to identify the majority of species.