Use of Diverse Extraction Protocols to Decide the Integrity of Deoxyribonucleic Acid (DNA) Samples Extracted from Bovine Bone Samples of Different Ages obtained from an Abattoir in Ikorodu, Lagos State, Nigeria

Abstract

Techniques for the identification human and non-human biological samples are developing at very high rates with the advent of different DNA extraction methods and polymerase chain reaction (PCR) based assays. This study aimed at using different extraction protocols to determine the integrity of DNA samples extracted from bovine bone samples of different ages collected from abattoir in Ikorodu, Lagos State, Nigeria. The DNA was extracted using CTAB, PCI protocols and a DNA kit (Quick DNA MiniPrep Plus Kit). Bovine mtDNA fragment containing the gene encoding ATPase 8 was amplified via Polymerase Chain Reaction (PCR). The PCR products were analysed on 1.8% agarose gel. It was observed that the DNA samples extracted using the PCI method at 48 h incubation time had the highest purity (1.68) and concentration (336 ng/µl) compared to other extraction methods employed in the study. However, DNA kit extracted samples had mean ± SE purity (1.52 ± 0-05) and concentration (192.25 ± 31.41 ng/µl) values that were higher than CTAB protocol values but lower than PCI protocol values.  All isolated DNA samples were PCR-worthy and thus yielded PCR products which were within the expected amplicon size of 126-bp. All DNA extraction protocols employed in this study are stable and efficient for use in the identification of aged non- human bones. This study also revealed that these protocols can be used to isolate PCR amplifiable DNA from old bones.