Comparative Account of DNA Extraction Protocols in Some Fresh Water Prawns of Genus Macrobrachium (Bate, 1868) (Family Palaemonidae) from Jammu Waters for PCR Based Applications
Raman Jasrotia*
and Seema LangerAnimal Cytogenetics Lab, Department of Zoology, University of Jammu, Jammu (180006), India.
Corresponding Author E-mail: ramanjasrotia18@gmail.com
Abstract: Genetic variations among prawns act as an important tool to characterize and differentiate between the species. Molecular and phylogenetic analysis of shrimps and prawns like any other organism rely on high yields of pure and better quality genomic DNA. In this regard isolation of DNA is the first and basic step. In spite of the availability of many protocols of DNA extraction from animal tissues, it is difficult to ascertain that which one would provide desired results for prawn tissue. In the present study, three different techniques of DNA isolation i.e., salting out, phenol-chloroform and Qiagen DNA extraction kit were performed and compared for their yield. Cephalothoracic tissue and muscle tissue of pleopods were used for isolation. Tissue samples from fresh specimens as well as from alcohol preserved specimens were employed for extraction. The quantity (µg/ml) and quality of isolated DNA were determined by UV spectrophotometry and agarose gel electrophoresis. Results showed that Phenol-chloroform method with slight modifications obtained higher yield of genomic DNA as compared to other methods. The present work also revealed that among fresh specimens cephalotoracic tissue yielded high concentration DNA than muscle tissue. However, among alcohol preserved specimens, the concentration of DNA was higher in muscle tissue of pleopods. The high quality DNA was then subjected to randomly amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) analysis. The DNAs produced clear, sharp and reproducible PCR (Polymerse chain reaction) product pattern.
Keywords: DNA Extraction; Genetic Variations; ISSR; PCR; Phenol-Chloroform; RAPD; Spectrophotometry Back to TOC