Standardization and Optimization of RAPD assay for genetic analysis of Noctuid species
Pawan Kumar, Shweta Thakur, V.K. Mattu and Raghav Dutta
The genomic DNA of various noctuid species belonging to the family Noctuidae has been subjected to RAPD-PCR analysis. The main objective under study was to standardize the methods to generate RAPD markers. Regarding standardization 100 bp ladder is used, amount of DNA template to be used is 100 ng/100 µl. Twenty random primers viz. OPA 1 to OPA 20 were employed to generate RAPD markers out of which OPA13 showed better results, different RAPD profiles were observed for the different species indicating that the RAPD-PCR analysis can be applied as a useful tool in quickly screening the strains to aid in discriminating these species. Finally, regarding the standardization of PCR product and loading dye the combination of 20 µl of PCR product with 4 µl of dye produced very refined bands with OPA 13. The potential for RAPD-PCR technique is to provide useful genetic data for the discrimination up to the inter-specific and intra-specific level of noctuid species is of concern and is hereby discussed.