An Efficient Method of Genomic DNA Extraction and Quantification from Insects

Abstract

The DNA extraction process is the most crucial  step for 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.