An Efficient Method of Genomic DNA Extraction and Quantification from Insects

Abstract

The DNA extraction process is the most crucial step demonstrating various molecular techniques such as PCR, DNA barcoding, metagenomics etc. This paper describes a simple and efficient protocol of DNA extraction from insect’s sample that included Honey bees, Carpenter bees and Flour beetle’s tissues with slight modification in CTAB method. DNA concentration together with purity has been determined by spectrophotometer absorbance reading at O.D260nm and O.D260/280nm respectively. Additionally, lambda DNA marker (NEB3012S) has been employed to validate band intensity by agarose gel electrophoresis. The DNA band exposed less smearing and intense band in agarose gel. Band of DNA in HB and CB lane was greater than 95.4ng but less than 95.4ng in lane FB as compared to lambda DNA marker bands. Also, DNA extracted from the tissue of Honey bee (HB), Carpenter bee (CB) and Flour Beetle (FB) measured by spectrophotometer for analysing O.D at 260 nm was found to be 4.9, 5.1 and 3.7ug/ml and the purity by considering 260/280 O.D ratio was 2.33, 1.76, 1.92. Absorbance ratio of 2.33 in DNA of Honey bees (HB) sample indicated slight contamination of protein or carbohydrate. Therefore, an achievable approach has been made to extract high quality of DNA by CTAB method and applying spectrophotometer for Optical Density as well as marker lambda DNA by agarose gel electrophoresis for quantity and quality analysis.