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A Simple Protocol to extract DNA from earthworm tissue for Molecular studies

Kushwaha T, Vishwakarma A, Rahul Paliwal, Sashidhar Burla and Shweta Yadav

A simple protocol has been developed to extract high quality of DNA from different species of earthworms of family Moniligastridae (Drawida travancorensis D.pellucida, D.parambikimala, D.modesta); Megascolecidae (Perionyx sansibaricus) Octochaetidae (Travoscolides chengannures); Lumbricidae (Octolasion cyaneum,Eisenia andrei, E.fetida) Eudrilidae (Eudrilus eugeniae) collected from western Ghats of India. A good quantity (261.6 to 151 μg/μl) of DNA was obtained. With present modified protocol. Ratio of A260/A280 was also in range of 1.91- 1.98 while, routine protocol was recorded 1.48-2.24 μg/μl. Extracted DNA was used to amplify 683 bp of the cytochrome oxidase I (COI) gene with LCO1490 and HCO2198 primers. All sequences of amplified gene were aligned, edited and analyzed using Maximum Likelihood method to characterize different species of earthworm. It appears that modified protocol used in present study was efficient and easy to extract good quality of DNA from different species of earthworms.