Study Area
Modern genetic techniques have simplified this mammoth task by means of DNA Barcodes, which are short sequences of mitochondrial and nuclear genes generally obtained by PCR (a powerful approach of molecular genetic analysis), which are subsequently used to identify and relate an individual at the species as well as higher taxonomic level.
In order to explore these barcodes, there is a need for universal primers, which can be used across wide range of species. Therefore Primer designing becomes a critical step and minor adjustments in the sequence can have large impact on barcode recovery. The present work emphasizes on designing universal primers (if feasible) or group specific primers, which would help in exploring anuran diversity through DNA barcoding analysis. For the purpose, efforts are made to identify Potential priming sites across a few mitochondrial as well as nuclear genes, viz., 12S/16S ribosomal RNA, CO1, CO2, CO3, Cytochrome-b, ND1, RAG1 and Rhodopsin, by comparative in-silico analysis of sequences available in the public domain.
The latter comprised of complete mitochondrial sequences of 27 anurans species representing archeobatrachia, mesobatrachia and neobatrachia, and reference nuclear sequences of >50 anurans. Primer sequences along with their location and utility are described. A preliminary validation study was carried with the primers designed for 12S ribosomal gene on different frog samples. These primers will also be instrumental in studying the evolutionary relationships among diverse anurans.
