Molecular identification of selected spiders by amplification of reference target DNA sequence and in silico analysis

Abstract

Spiders or Araneae is one of noteworthy groups of carnivorous invertebrates that provide important model systems for study of ecosystem in the world. Molecular approach was conducted to characterize spider species as the identification of spiders through morphology is often challenging. Accurate morphological characterization of spider species is always time consuming and only practicable for a particular life stage or gender of the specimen. In this research, molecular identification of selected spiders was demonstrated by amplification of reference target DNA sequences based on primer pairs used and subsequently in silico analysis. A total of three selected spider species were used for DNA extraction by using TriZOLTM extraction protocol. Amplification of reference target region of mitochondrial DNA by using specific primers followed by DNA sequencing and phylogenetic analysis were carried out. This research also provided evidence that TriZOLTM extraction protocol is effective for producing good quality of DNA from the samples. The resolving potential of specific primer pairs that amplify the 660 bp of mitochondrial Cytochrome C Oxidase Subunit 1 (CO1) gene is also proved to be able to discriminate between spider species. The length of Polymerase Chain Reaction (PCR) products amplified is ranged from 651 bp to 704 bp for all the tested samples. The average high genetic similarity after comparison of database in NCBI though Basic Local Alignment Tool Search (BLAST) revealed that the analysed individuals that involved in this research were placed in the correct species, although the profile was based on only a single representative of each species. Phylogenetic analysis towards the reference target DNA sequences consequently provides unique insight into the evolutionary relationship of each analysed samples.