Abstract
Microbial biotechnology is a continually expanding research area with significant economic value and applications in diverse areas including pharmaceuticals, molecular biology, industrial processes, biocatalysis, medicine and bioremediation. The number of bacterial species which have been isolated, characterized and sequenced continues to expand at an ever increasing rate.
Researchers aiming to isolate novel bacterial strains from environmental samples face multiple challenges including identifying clonality of cultures and classifying bacterial strains. Traditionally, sequencing of 16SrRNA gene segments has been used for such purposes; oftentimes this requires designing multiple primer sets to enable amplification of target sequences across abroad range of bacteria. Successfully sequencing 16SrRNA genes from uncharacterized organisms therefore presents a challenge.
DArTseq genotyping by sequencing technology provides an alternative method for characterizing bacterial isolates based on sequence data in an affordable and high-throughput manner, without reliance on prior sequence information for primer design. DArTseq genomic representations are produced through simple and reproducible restriction enzyme and PCR based protocols, allowing samples to be processed highly quantitatively, quickly and cheaply. Here we demonstrate the application of DArTseq for identification of thermophilic microorganisms isolated from environmental samples.