DNA PROFILING: IDENTIFYING SPECIES, INDIVIDUAL AND SEX OF COMMON LEOPARD

For effective conservation of species, it is always very important to ascertain species, sex and genetic structure in the wild crime cases. These provide additional insight in understanding biology of the species across its geographical region. Therefore, there was a need to establish appropriate protocols for DNA extraction, species and sex identification. Present book mainly focuses on DNA extraction, species & sex identification and assigning genetic diversity in leopards (Panthera pardus). Different methods used for the DNA extraction, Qiagen kit based protocol for skin DNA isolation and the quality (>500 bp to >1 kb) & quantity was good. Further mitochondrial DNA partial fragment of 12s rRNA and 16s rRNA genes were tested for species identification along with different bioinformatics tools like sequencing analysis, BLAST, FASTA, Multiple Sequence Alignment (MSA), SEQUENCING ANALYSIS 5.2, BIOEDIT 7.0.9.0, SEQUENCHER 4.7 etc. Sex identification was carried out by using several combinations of primers for sex specific genes. Sex was confirmed after cross checking the results of these combinations on the basis of presence and absence of specific amplification. Of the eight skins examined, 3 were of male whereas remaining 4 were of female. Multilocus genotyping was also conducted to identify the individuals using the multiplex PCR with different combinations of primers for microsatellite loci which were initially developed for Bengal tiger (Panthera tigris tigris), domestic cat (Felis catus), and snow leopard (Uncia uncia). This book provides the overall view of molecular techniques to use in wildlife forensics and conservation sciences for effective conservation and management of wildlife species.