Pages: 01-06

Extraction of genomic DNA from hemolymph of insects by an efficient and quick method

Author: M. Kannan, N. K. Singh, P. Britocathrin, S. Shantkriti, B. Padmanaban and M. Krishnan

Category: Biology

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Abstract:

An extraction of high quality genomic DNA is the first and crucial step in molecular biology experiments for the study of molecular taxonomy, diversity and evolution. In general, traditional methods and kits for the extraction of genomic DNA from any biological sample are cost effective but time consuming and use toxic chemicals. The present study described a simple method for the extraction of genomic DNA from haemolymph of insects using Haemocyte lysis buffer (HLB). HLB method is quicker as it has few steps, and doesn't use toxic chemicals. HLB method yielded approximately 60 -120 g of total genomic DNA from haemolymph per isolate (50 L) from the selected insects and the purity of genomic DNA ranged between 1.61 - 1.83 at 260 / 280 nm as revealed by spectrophotometry analysis. The quantity and quality of genomic DNA was compared with kit methods key. The electrophoretic analysis of the genomic DNA revealed nucleotides above 10 kb without protein and RNA contamination. Absence of inhibitory compounds from the extracted DNA was confirmed by amplification of β-actin and Cytochrome oxidase subunit 1 genes and restriction digestion analysis using EcoR1 enzyme. The overall results suggested that the extraction of genomic DNA through this method is easy and appropriate for molecular biology experiments.

Keywords: Insect haemolymph, Haemocyte lysis buffer, genomic DNA, ?-actin, cytochrome oxidase 1, restriction digestion


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